LOCUS       V036529                 7580 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036529
VERSION     V036529
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..3100
                     /label="Mark2(NM_001080389)"
                     /note="Mark2(NM_001080389)"
                     /gene="Mark2"
     polyA_signal    3147..3371
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3417..3845
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3859..4188
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4255..5049
                     /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    5223..5344
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5393..5409)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5417..5433
                     /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(5441..5471)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5486..5507
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5795..6380)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6551..7411)
                     /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(7412..7516)
                     /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 tccagcgctc ggacccccct acccacgctg aacgaaaggg acacggagca
      961 gcccaccttg ggacaccttg attccaagcc cagcagtaag tccaacatgc tgcggggccg
     1021 caactcagcc acctctgctg acgagcagcc ccatattggc aactaccggc tccttaagac
     1081 cattggcaag ggtaactttg ccaaggtgaa gttggcccgg cacatcctga cggggaaaga
     1141 ggtagctgtg aagatcatcg acaagaccca gctgaactcc tccagcctac agaaactgtt
     1201 ccgagaagta agaataatga aggttttgaa tcatcccaac atagttaagt tgtttgaagt
     1261 gatcgagact gagaagactc tctaccttgt catggagtat gccagtggcg gagaggtgtt
     1321 tgattaccta gtggcccatg gcaggatgaa agaaaaagaa gctcgagcca aatttcgcca
     1381 gatagtgtct gctgtgcagt actgtcacca gaagttcatt gttcatagag atctaaaggc
     1441 agaaaacctg ctcctggatg ctgatatgaa catcaagatt gcagactttg gctttagcaa
     1501 cgaattcacc tttgggaaca agctggatac tttctgtggc agtcctcctt atgctgcccc
     1561 agaacttttc cagggcaaaa agtatgatgg tcctgaggtg gatgtctgga gcctgggagt
     1621 catcctctat acactggtca gcggatccct gccttttgat ggacagaacc tcaaggagct
     1681 gcgggaacgg gtactgaggg ggaaataccg tattccgttc tacatgtcca cggactgtga
     1741 aaatctgctt aagaaatttc tcatacttaa tcctagtaag agaggcactt tagagcaaat
     1801 tatgaaagat cggtggatga acgtggggca tgaggacgat gagctaaagc cttatgtgga
     1861 acctctccct gactacaagg acccccggcg gacagagttg atggtgtcaa tgggttacac
     1921 acgggaagag atccaggact cgctggtagg ccagaggtac aacgaagtga tggctaccta
     1981 tctgctcctt ggctacaaga gctctgagct ggaaggtgat accatcactt tgaagccccg
     2041 gccttcagct gatctaacca acagcagtgc cccatctcca tcccacaagg ttcagcgcag
     2101 cgtctctgcc aaccccaagc aacgacgctc cagtgaccag gccgtccctg ccattcccac
     2161 ctcgaattcc tactctaaga agactcagag taacaacgca gaaaataagc ggcctgagga
     2221 agagacaggg cggaaagcca gcagcaccgc caaagtgcct gccagccctc tgcctggcct
     2281 ggacaggaag aagaccactc ctgccccctc cacgaacagc gtcctttcca ccagcacaaa
     2341 ccgaagcagg aactccccac ttttggacag ggccagcctt ggccaggcct ccatccagaa
     2401 tggtaaagac agcacagccc cccagcgcgt ccctgtcgcc tccccctccg cccacaacat
     2461 cagcagcagt agtggagccc cagaccgaac taatttccca cggggtgtgt ccagtcgaag
     2521 caccttccat gctgggcagc tccgacaggt gcgggaccag cagaatctac cctacggtgt
     2581 gaccccagcc tctccctctg gccatagcca gggccggcgg ggggcctctg gcagcatctt
     2641 cagcaagttc acctccaagt ttgtccgcag aaatctgtct ttcaggtttg ccagaaggaa
     2701 cctgaatgaa cctgaaagca aagaccgagt ggagacgctc agacctcacg tggtaggcag
     2761 tggaggcact gacaaggaca aggaggagtt tcgggaggcc aagcctcgct ccctgcgctt
     2821 cacctggagc atgaagacca cgagctctat ggagcccaat gagatgatgc gggagatccg
     2881 caaggtgctg gacgccaaca gctgccaaag cgagctgcac gagcggtaca tgctactgtg
     2941 cgtgcatggc acaccaggcc acgagaactt tgtgcagtgg gagatggagg tgtgcaaact
     3001 gccccggctg tctctcaacg gtgttcggtt taagcggata tcgggcactt ccatggcctt
     3061 caaaaacatt gcctccaaaa tagccaatga gctgaagctt taactcgagt ctagagggcc
     3121 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3181 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3241 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3301 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3361 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     3421 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3481 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3541 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3601 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3661 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3721 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3781 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3841 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3901 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3961 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4021 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4081 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     4141 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4201 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4261 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4321 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4381 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4441 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4501 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4561 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4621 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4681 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4741 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4801 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4861 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4921 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4981 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5041 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5101 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5161 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5221 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5281 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5341 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5401 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5461 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5521 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5581 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5641 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5701 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5761 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5821 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5881 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5941 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6001 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6061 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6121 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6181 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6241 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6301 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc
     6361 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6421 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6481 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6541 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6601 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6661 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6721 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6781 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6841 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6901 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6961 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7021 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7081 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7141 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7201 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7261 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7321 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7381 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7441 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7501 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     7561 atctcccgat cccctatggt
//
