LOCUS       V029980                 8139 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029980
VERSION     V029980
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2819
                     /label="Siglecg(NM_172900)"
                     /note="Siglecg(NM_172900)"
                     /gene="Siglecg"
     CDS             2826..2891
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2915..3503
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3569..3617
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3819..4247
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4261..4590
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4657..5451
                     /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    5627..5748
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5797..5813)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5821..5837
                     /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(5845..5875)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5890..5911
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6199..6787)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6958..7818)
                     /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(7819..7923)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgtc actgctgctg ttcctgctgt
      781 ccttcctgtt ggatgggcct caaggtcaga tggagagcta cttcctacag gtgcagagaa
      841 ttgtgaaggc acaggagggt ctgtgcatct tcgtgccctg ctccttctcc tcccctgagg
      901 gaaaatggct taaccgttcc ccactttatg gctactggtt caaaggcatc agaaaaccat
      961 cactttcatt tccagtggcc acaaataaca aagataaagt gttagaatgg gaagcccgag
     1021 ggcgcttcca gctcttgggg gatatctcta aaaagaactg ttccttgcta atcaaagatg
     1081 ttcagtgggg agactcaaca aactatttct tccggatgga gagaggattt gagagattca
     1141 gtttcaagga ggagttcagg ctacaagtgg aagccctgac tcagaagcca gatatcttca
     1201 ttcctgaggt cctggagcct ggggagccag tgaccgttgt ctgcttgttt tcctggacct
     1261 tcaaccaatg cccagctcct tctttctcct ggatggggga tgctgtctcc ttccaagaaa
     1321 gcagaccgca cacatccaat tactcagttc tgagctttat cccaggactt caacaccatg
     1381 atactgagct cacatgtcag ctggacttct ctagaatgag cacacaaagg actgtccgac
     1441 taagagtggc ctatgccccc agatctcttg ctatcagcat tttccatgac aatgtatcag
     1501 tcccagactt gcatgagaat ccttcacatc tggaagttca gcaaggccag tctcttcgcc
     1561 tcctctgtac tgctgacagc cagccccctg ctacactgag ctgggtccta gaggaccaag
     1621 tcctttcttg gtctagccct gtggggtcga gaaccctggc actagagctg ccctgggtga
     1681 aagctgggga ctctggacac tacacttgcc aagcagagaa taggctgggc tcccagcaac
     1741 ataccctgga cctctctgtg ctgtaccccc cacaggacct gagagtgact gtttcccaag
     1801 caaacaggac agtgttggaa atcctcagga atgccatctc cctcccagtc ctggagggcc
     1861 aaagcctgtg cctagtctgt gtcacctata gcaatccccc agccaatgtg agttgggctt
     1921 gggtgacaca gaccctgatc ccaatccagt cttcagagcc tggggtcctg gagctgcctc
     1981 tggtccagag agaacatgaa ggagaattca cctgtgctgc acagaaccca ctgggtgccc
     2041 agcgcatctc tctgagcctc tctgtgcact acccgcccca gatgtccagc ccctcctgct
     2101 cctgggaggc caagggtctg cactgcaact gctcctccag agcctggcct gccccctccc
     2161 tgcgctggcg gctgggggag gggctgctgg aggggaacag cagcaatgcc tccttcacag
     2221 tcactttcag ctcacttgga ccctgggtca acagctccct gagcctcctt caggagctgg
     2281 ggcccagcct ctggctcagc tgtgagtcct ggaacaccca tggagcccag accacctctg
     2341 tcctgctgct acctgataaa gacagtgcca cagcattctc caagggagca gttttgggct
     2401 ttggcatcac agccctcctt gccctgtgcc tcatcgtggt catcgtgaag accctccaga
     2461 agaaaggaac ccaggaggaa ccctccaggc ccaagctctc ccggggcagc acgatcctgg
     2521 actacatcaa tgtggtcccc aagaccaggt ccctggctcg gaattggaaa gctgaaccag
     2581 atgccccttc taggagctca cccctggaca ctcacttccc taaaccaaag aagaagcaaa
     2641 aggacccaca tttcacgtac cctggttgcc cagatcccac atcatcctct caggtcccag
     2701 tctctgagaa taacccggaa gaactccact atgctgctct caacttctcc cgcctgagac
     2761 tgcaggagac ccaggatccc caagacacct attctgatta cacagaggtc agagtccacg
     2821 gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     2881 acgatgacaa gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg
     2941 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     3001 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     3061 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     3121 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     3181 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     3241 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     3301 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     3361 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     3421 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     3481 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag
     3541 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg
     3601 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     3661 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     3721 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     3781 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3841 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3901 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3961 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4021 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4081 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4141 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     4201 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     4261 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4321 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4381 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4441 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4501 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4561 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4621 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4681 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4741 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4801 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4861 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4921 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4981 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5041 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5101 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5161 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     5221 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5281 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5341 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5401 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5461 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5521 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5581 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5641 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5701 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5761 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5821 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5881 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5941 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6001 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6061 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6121 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6181 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6241 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6301 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6361 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     6421 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6481 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6541 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6601 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     6661 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6721 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6781 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6841 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6901 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6961 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7021 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7081 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7141 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7201 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7261 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7321 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7381 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7441 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7501 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7561 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7621 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7681 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7741 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7801 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7861 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7921 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     7981 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     8041 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     8101 caaggcttga ccgacaattg catgaagaat ctgcttagg
//
