LOCUS       V041656                10511 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041656
VERSION     V041656
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             951..5342
                     /label="CECR2(NM_001290047)"
                     /note="CECR2(NM_001290047)"
                     /gene="CECR2"
     CDS             5349..6044
                     /codon_start=1
                     /note=""
                     /product="promiscuous R40G mutant of a biotin protein
                     ligase from Aquifex aeolicus (Kim et al., 2016)"
                     /transl_table=1
                     /translation="FKNLIWLKEVDSTQERLKEWNVSYGTALVADRQTKGRGGLGRKWL
                     SQEGGLYFSFLLNPKEFENLLQLPLVLGLSVSEALEEITEIPFSLKWPNDVYFQEKKVS
                     GVLCELSKDKLIVGIGINVNQREIPEEIKDRATTLYEITGKDWDRKEVLLKVLKRISEN
                     LKKFKEKSFKEFKGKIESKMLYLGEEVKLLGEGKITGKLVGLSEKGGALILTEEGIKEI
                     LSGEFSLRRS"
                     /label="BioID2"
     CDS             6045..6071
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    6107..6331
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      6377..6805
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6819..7149
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             7216..8010
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    8184..8305
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(8354..8370)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8378..8394
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(8402..8432)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8447..8468
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8756..9344)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9515..10375)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(10376..10480)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gcttaaggcc tgttaaccgg tcgtacgtct ccggattcga attcgccacc atgtgcccag
      961 aggagggcgg cgcggccggg ctgggcgagc tccgctcctg gtgggaggtc ccggccatcg
     1021 cgcacttctg ctcgctcttt cgcaccgcgt tccgcctgcc cgacttcgag atcgaggagt
     1081 tagaagccgc tcttcacaga gatgacgtgg agtttatcag tgacctgatt gcctgcctgc
     1141 ttcagggctg ctatcaacga agagatatca cgcctcagac attccacagc tacctagagg
     1201 acatcatcaa ctaccgctgg gagctcgaag aagggaagcc caaccctctg agggaagcca
     1261 gtttccagga cctgcctctt cgcacacggg tggagatcct gcaccgactc tgtgattacc
     1321 ggctggatgc agacgatgtc ttcgatcttc taaagggcct ggatgcagac agtctccgtg
     1381 tggagccatt gggtgaagac aattctgggg cactatattg gtatttctat ggaacacgaa
     1441 tgtacaaaga ggacccggtg caaggaaaat ccaatggaga actctctttg agcagggaaa
     1501 gtgaaggaca aaaaaatgtc tcaagtattc ctggaaaaac gggaaaaaga agaggaagac
     1561 ccccaaaacg gaagaaactg caggaggaga ttctgttgag tgaaaagcag gaagaaaatt
     1621 ccttggcatc cgagccacag acaagacatg ggtcccaagg gccaggccaa ggtacttggt
     1681 ggctcctgtg ccagacagaa gaggaatgga gacaggtcac cgagagtttt cgcgagagga
     1741 cctcccttcg agaacggcag ctctacaagc tcctcagtga ggacttcctg cctgagatct
     1801 gcaacatgat cgcccagaag ggaaaacgtc cacagcgcac aaaggcagag ttgcatccta
     1861 ggtggatgtc tgaccacctg tccatcaaac ccgtcaagca agaggagact cctgtgctga
     1921 ccagaataga aaaacaaaag cgcaaagagg aggaagaaga gcgtcagatt cttctagcag
     1981 tgcagaagaa ggagcaggag cagatgctaa aggaagagag gaaacgcgag ttggaggaga
     2041 aggtcaaggc agtggaagat cgagcgaaga ggagaaagct cagggaagaa agggcatggc
     2101 tgctggctca aggaaaggag ctccctccag aactttccca tctggacccc aattccccca
     2161 tgagagagga aaaaaagact aaagacctct ttgagttgga tgatgatttc actgctatgt
     2221 ataaagttct agacgtggta aaggctcaca aggattcctg gcccttcttg gaacctgtgg
     2281 atgaatctta tgcccctaac tattatcaga ttattaaggc ccccatggat atttccagca
     2341 tggagaagaa actgaatgga ggtttatact gtaccaagga ggaatttgta aatgacatga
     2401 agaccatgtt caggaattgt cgaaagtata atggggaaag tagtgagtat accaagatgt
     2461 ctgataattt agagaggtgt ttccatcggg caatgatgaa acattttcct ggagaagatg
     2521 gagacacaga tgaagaattt tggattcgag aggatgaaaa gcgggagaaa agacggagtc
     2581 gggctgggcg aagtggtggg agccatgttt ggacccgctc cagggaccca gaagggtcca
     2641 gcaggaaaca gcagcccatg gagaatggag gaaagtcgtt gccccccaca cgccgagcgc
     2701 cctcttctgg ggacgatcag agcagcagct ccacacagcc cccgcgggag gtgggcactt
     2761 ccaatggccg aggtttttct catcccctgc attgtggtgg gacacccagc caggcaccct
     2821 ttttaaacca gatgaggcca gcagtaccag gaacatttgg ccctctgcga ggatcagatc
     2881 ctgccacctt gtatggctcc tctggagtcc cggagccaca ccccggggag cctgtgcagc
     2941 agcgtcagcc tttcaccatg cagcctccag ttggaattaa cagcctccga ggacccaggc
     3001 taggcacacc agaggagaag caaatgtgcg gggggctgac acacctttct aacatgggcc
     3061 cacaccctgg atccttgcag cttgggcaga taagtggccc aagtcaggat ggaagcatgt
     3121 atgctccagc tcagttccag ccaggattca ttcctccccg gcatgggggg gctccagccc
     3181 ggccaccaga ctttcctgaa agctcagaaa ttcctcccag ccatatgtat cgatcgtaca
     3241 agtacctgaa tcgagtacac tctgccgtct ggaatgggaa ccatggtgct acgaaccaag
     3301 gacccttggg cccagatgag aagccccacc tggggccagg accctctcac cagcctcgca
     3361 ctctcggtca cgtgatggat tcccgagtca tgagaccacc tgtccccccc aaccagtgga
     3421 ctgaacaatc aggcttccta cctcatggag ttccttcctc agggtacatg cgaccgccct
     3481 gcaagtctgc cggacatcgg ttacagccac ctccagtgcc agcacccagt tctttgtttg
     3541 gagcacctgc ccaggctctt cggggggtgc agggagggga ctccatgatg gacagcccag
     3601 agatgattgc gatgcagcag ctctcctccc gcgtctgccc cccaggtgtg ccttaccacc
     3661 cccaccagcc tgcacacccc cgtttacctg gcccttttcc gcaggtagct cacccaatgt
     3721 cagtcactgt gtcagccccc aagcctgccc tgggcaaccc tgggagggca ccggagaaca
     3781 gtgaagcaca agagcctgag aatgaccaag cagagccgtt gcctggcctt gaagagaaac
     3841 caccaggtgt tggtacttca gagggggtct acctcacaca actacctcac cccacacctc
     3901 ccctgcagac tgactgcacc aggcagagct caccacaaga aagggaaaca gtgggcccgg
     3961 agctcaaaag cagctcctcc gaatctgcgg acaactgtaa agcaatgaag ggcaagaatc
     4021 cctggccctc ggatagcagc taccccggcc cagccgccca agggtgcgtg agagacctct
     4081 ccacggtggc agacaggggc gctctatccg agaacggagt cattggggaa gcatctcctt
     4141 gtggatcgga ggggaagggc cttggtagca gtggttccga aaagctgctc tgccccagag
     4201 gcagaacgtt gcaggaaacc atgccatgca cgggacagaa cgcagcgaca ccgcccagca
     4261 cagaccccgg tttgacggga ggcactgtga gccagtttcc cccgctgtat atgcctggcc
     4321 tagagtaccc gaattcagct gcccattacc acatcagtcc aggcctgcag ggtgtgggcc
     4381 ctgtgatggg agggaagtcc ccagcatccc atccccagca ttttccccca aggggctttc
     4441 agtctaacca cccacattct ggaggctttc cccggtatcg ccccccacaa ggaatgaggt
     4501 attcctacca cccaccgcca cagccttcct accaccacta tcagcgaact ccttactatg
     4561 cctgtccaca gagcttttct gactggcaga gacctctcca tccccaggga agcccaagcg
     4621 gacccccagc cagtcagcct cccccaccaa ggtccctctt ctcagataag aatgccatgg
     4681 ccagtctgca aggctgtgag acactgaatg ctgccttaac ttctccaacc cgtatggatg
     4741 cagtggctgc taaagtccca aatgacgggc agaatcctgg tccagaggaa gagaagctgg
     4801 atgaatctat ggagaggcca gagagtccca aagaattttt agacctggac aaccataacg
     4861 cagctaccaa gcggcagagc tcgttgtcag ccagcgagta tctctatgga actcctccgc
     4921 ctctgagttc aggaatggga tttggttcat ctgcatttcc accccacagt gtgatgctgc
     4981 agacggggcc tccctatacc cctcagcggc cggccagtca ctttcagccc agggcttact
     5041 cttcccctgt ggctgccctc ccacctcacc acccaggggc cacccagccc aacggcctct
     5101 ctcaggaggg tcccatctat cgctgccagg aagaaggcct gggtcacttt caagctgtga
     5161 tgatggaaca aattggcact agaagtggaa taagaggacc tttccaggaa atgtacagac
     5221 catcaggaat gcagatgcac ccggtccagt cgcaggcctc gttcccaaag acccccacag
     5281 cagcaacatc acaggaggag gtgccgcctc ataagcctcc aacacttccc ctggatcaga
     5341 gcggatcctt caagaacctg atctggctga aggaggtgga cagcacccag gagagactga
     5401 aggagtggaa cgtgagctac ggcaccgccc tggtggccga cagacagacc aagggcagag
     5461 gcggcctggg cagaaagtgg ctgagccagg agggcggcct gtacttcagc ttcctgctga
     5521 accccaagga gttcgagaac ctgctgcagc tgcccctggt gctgggcctg agcgtgagcg
     5581 aggccctgga ggagatcacc gagatcccct tcagcctgaa gtggcccaac gacgtgtact
     5641 tccaggagaa gaaggtgagc ggcgtgctgt gcgagctgag caaggacaag ctgatcgtgg
     5701 gcatcggcat caacgtgaac cagagagaga tccccgagga gatcaaggac agagccacca
     5761 ccctgtacga gatcaccggc aaggactggg acagaaagga ggtgctgctg aaggtgctga
     5821 agagaatcag cgagaacctg aagaagttca aggagaagag cttcaaggag ttcaagggca
     5881 agatcgagag caagatgctg tacctgggcg aggaggtgaa gctgctgggc gagggcaaga
     5941 tcaccggcaa gctggtgggc ctgagcgaga agggcggcgc cctgatcctg accgaggagg
     6001 gcatcaagga gatcctgagc ggcgagttca gcctgagaag aagctaccct tacgatgtac
     6061 cggattacgc atagaagctt aagtttaaac cgctgatcag cctcgactgt gccttctagt
     6121 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     6181 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     6241 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     6301 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     6361 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     6421 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     6481 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct
     6541 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     6601 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     6661 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     6721 tattcttttg atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg
     6781 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     6841 agtccccagg ctccccaggc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     6901 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     6961 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     7021 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     7081 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     7141 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     7201 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     7261 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     7321 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     7381 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     7441 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     7501 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     7561 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     7621 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     7681 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     7741 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     7801 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     7861 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     7921 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     7981 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     8041 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     8101 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     8161 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     8221 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     8281 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     8341 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     8401 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     8461 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     8521 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     8581 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     8641 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     8701 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     8761 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     8821 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     8881 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     8941 cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     9001 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     9061 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     9121 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     9181 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     9241 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     9301 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     9361 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     9421 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     9481 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     9541 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     9601 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     9661 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     9721 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     9781 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     9841 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     9901 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     9961 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
    10021 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
    10081 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
    10141 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
    10201 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
    10261 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
    10321 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
    10381 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
    10441 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
    10501 cacctgacgt c
//
