LOCUS       V040258                 7520 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040258
VERSION     V040258
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             908..2926
                     /label="Dpysl2(NM_001378767)"
                     /note="Dpysl2(NM_001378767)"
                     /gene="Dpysl2"
     CDS             2963..3052
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    3087..3311
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3357..3785
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3799..4128
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4195..4989
                     /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    5163..5284
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5333..5349)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5357..5373
                     /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(5381..5411)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5426..5447
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5735..6320)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6491..7351)
                     /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(7352..7456)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg gccgagagaa agcaatctgg aaaggcagca gaggacgaag aggtccctgc
      961 cttttttaaa aacctgggct ccggcagtcc caagccgcgg cagaaattct gcggcatgtt
     1021 ctgccctgtg gaagggtcct cggagaacaa gaccatcgac ttcgactcgc tttcagtggg
     1081 ccggggctcg gggcaagtgg tggctcagca gcgcgatgtc gcccacttgg gccccgaccc
     1141 gcaccggcag agccggcgtg ccagtgggga gccatctgtt gaatcgagcc ggaaggtgga
     1201 gatccggcgg gcctcgggca aggaagccct gcagaacatc tatgatcaga gcgatcgtct
     1261 tctgatcaaa ggtggcaaga ttgtgaatga tgaccagtcc ttctatgcag acatatacat
     1321 ggaagatggg ttgatcaagc aaataggaga aaacctgatt gtaccaggag gggtgaagac
     1381 catcgaagcc cactccagaa tggtgattcc cggagggatt gacgtgcata ctcgcttcca
     1441 gatgcctgac cagggaatga catccgctga tgacttcttc cagggaacca aggcggccct
     1501 ggccggggga accaccatga tcattgacca tgttgttcct gagcccggga cgagcctatt
     1561 ggctgccttt gatcagtgga gggagtgggc tgacagcaag tcctgctgtg actattcgct
     1621 gcacgtggac atcactgagt ggcacaaggg catccaggag gagatggaag ctctggtgaa
     1681 ggaccacggg gtaaactcct tcctcgtgta catggctttc aaagatcgat tccagctgac
     1741 ggattcccag atctatgaag tgctgagcgt gatccgggat atcggtgcca tagctcaagt
     1801 ccacgcagag aatggtgaca tcattgctga ggaacagcag aggatcctgg atctgggcat
     1861 cacgggcccc gagggacacg tgttgagccg gccagaggag gtcgaggctg aagctgtgaa
     1921 ccggtccatc actattgcca accagaccaa ctgccctctg tatgtcacca aagtgatgag
     1981 caagagtgcg gctgaagtca tcgctcaggc acggaagaag ggaactgtgg tgtatggtga
     2041 gcccatcacg gccagcctgg ggactgatgg ctctcattac tggagcaaga actgggccaa
     2101 ggctgcggcc tttgtcacct ccccaccctt gagccccgac ccaaccactc cagactttct
     2161 caactcgttg ctgtcctgtg gagacctcca agtcactggc agtgcccact gcaccttcaa
     2221 cactgcccag aaggctgtgg ggaaggacaa cttcaccttg attcccgagg gcaccaacgg
     2281 cactgaggag cggatgtctg tcatttggga taaagctgtg gtcactggga agatggatga
     2341 gaatcagttt gtggctgtga ccagcaccaa cgcagccaaa gtcttcaacc tttacccccg
     2401 gaaaggtcgc atctcggtgg gatctgatgc tgacttggtc atctgggacc ctgacagtgt
     2461 gaagaccatc tctgccaaga cacacaacag tgctcttgag tacaacatct ttgaaggcat
     2521 ggagtgtcgc ggctccccac tggtggtcat cagccagggc aagattgtcc tggaggacgg
     2581 cacacttcat gtcactgaag gctcaggacg ctacattccc cggaagccct tccctgactt
     2641 tgtgtacaaa cgcatcaaag caaggagcag gctggctgag ctgagagggg tccctcgtgg
     2701 cctgtatgac ggaccggtat gcgaggtgtc tgtgacgccc aagacggtga ctccagcctc
     2761 atcagctaag acatcccctg ccaagcagca ggcaccacct gttcggaacc tgcaccagtc
     2821 tggattcagc ttgtctggtg ctcagattga cgacaacatt ccccgccgca ccacccagcg
     2881 catcgtggca ccccctggtg gccgtgccaa catcaccagc ctgggcggtg gaggcggatc
     2941 aggaggcggt ggatctctcg aggaacaaaa actaatatca gaggaagatt tggaacagaa
     3001 actaatcagt gaagaagatt tagagcaaaa gttaatttcc gaggaggact tatgagggcc
     3061 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3121 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3181 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3241 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3301 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     3361 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3421 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3481 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3541 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3601 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3661 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3721 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3781 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3841 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3901 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3961 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4021 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     4081 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4141 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4201 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4261 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4321 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4381 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4441 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4501 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4561 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4621 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4681 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4741 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4801 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4861 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4921 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4981 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5041 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5101 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5161 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5221 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5281 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5341 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5401 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5461 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5521 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5581 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5641 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5701 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5761 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5821 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5881 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5941 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6001 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6061 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6121 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6181 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6241 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc
     6301 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6361 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6421 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6481 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6541 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6601 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6661 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6721 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6781 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6841 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6901 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6961 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7021 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7081 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7141 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7201 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7261 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7321 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7381 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7441 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     7501 atctcccgat cccctatggt
//
