LOCUS       V034141                 9142 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034141
VERSION     V034141
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             884..1591
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             1628..4645
                     /label="MIB1(NM_020774)"
                     /note="MIB1(NM_020774)"
                     /gene="MIB1"
     CDS             4646..4663
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    4709..4933
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4979..5407
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5421..5750
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5817..6611
                     /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    6785..6906
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6955..6971)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6979..6995
                     /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(7003..7033)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7048..7069
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7357..7942)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8113..8973)
                     /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(8974..9078)
                     /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 ggctagcgct accggtcgcc accatggtga gcaagggcga
      901 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
      961 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     1021 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     1081 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     1141 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     1201 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     1261 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     1321 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     1381 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     1441 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     1501 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     1561 ctccaccggc ggcatggacg agctgtacaa gggtggaggc ggatcaggag gcggtggatc
     1621 tctcgagatg agtaactccc ggaataaccg ggtgatggtg gaaggggttg gcgctcgggt
     1681 agtgcgcggc ccggactgga agtgggggaa gcaggacggc ggcgagggcc atgtgggcac
     1741 cgtccggagc ttcgagagcc ccgaggaggt ggtggtagtg tgggacaacg gcacagctgc
     1801 caactaccgc tgctccgggg cttacgacct ccgcatcctg gacagcgcgc ccaccggcat
     1861 caagcatgat ggaaccatgt gtgatacctg ccgccagcaa ccaatcattg gcattcgatg
     1921 gaagtgtgca gagtgtacaa attatgattt gtgcacagtg tgttatcatg gagataaaca
     1981 tcatttaaga catcgctttt accgaattac tacaccggga agtgagaggg ttctgttaga
     2041 gtctcgtagg aaatctaaga agattacagc cagaggaatc tttgcaggtg ccagagtggt
     2101 gcgaggagtg gactggcagt gggaagatca agatggagga aatggacgta ggggaaaggt
     2161 aacagaaatc caggactgga gtgcatcaag cccacatagc gcagcatatg tcctctggga
     2221 taatggtgct aagaaccttt acagagttgg ctttgagggc atgtctgatc tgaaatgtgt
     2281 ccaggatgcc aagggaggtt ctttctacag agatcactgc cctgtgctag gtgagcagaa
     2341 tggcaacagg aatcctggtg gattgcagat tggtgacctg gtaaatatag atctcgacct
     2401 cgaaattgta cagtctttgc agcatggtca tggaggatgg actgatggaa tgtttgagac
     2461 tttaactaca actggaactg tttgtggcat tgatgaagat catgacattg tagtacagta
     2521 tccaagtggc aataggtgga ccttcaatcc tgctgttctc actaaagcga acattgtccg
     2581 aagtggagat gctgctcagg gtgcagaagg aggcacctcg cagtttcaag tgggtgatct
     2641 tgtacaagtt tgttatgacc tggaacgaat taaacttcta caaagaggac atggagaatg
     2701 ggctgaagcg atgcttccaa ctttaggtaa agttggccga gtacaacaga tttattcaga
     2761 cagtgattta aaggtggaag tttgtggaac atcttggaca tacaatccag cagcagtttc
     2821 caaggtggca tctgcaggat cagccattag caatgcatct ggtgaaagac tctcacaact
     2881 cctgaagaaa ttatttgaaa cccaagaatc tggtgacctc aatgaagaat tagttaaggc
     2941 tgctgccaat ggagatgttg ctaaagtgga agatttgctt aaaagaccag atgtggatgt
     3001 aaatgggcaa tgtgctggcc acacagctat gcaagctgct agtcagaatg gacatgttga
     3061 cattttgaag ttacttttga agcaaaacgt ggatgtcgaa gcagaggata aagatggtga
     3121 tagagcagtt caccatgcag cttttggaga tgaaggcgct gttatagaag tactacatcg
     3181 aggtagtgct gatttgaatg ctcgaaacaa gcgccgacag acaccacttc atattgctgt
     3241 caataaaggt catcttcaag ttgtgaagac tttattggac tttggctgtc atcccagtct
     3301 ccaggattct gaaggtgata cccctcttca tgatgcaata agtaagaaac gtgatgatat
     3361 cctagcagtt cttttggaag ctggagcaga tgttaccatc acaaacaata atggatttaa
     3421 tgctctgcat catgctgcac taaggggaaa tcccagtgca atgcgtgttt tactatctaa
     3481 attaccaaga ccatggattg tggatgagaa gaaagatgat ggttatactg ccttacatct
     3541 ggctgccctt aataatcacg tagaagtggc tgaactgttg gtacatcagg gtaatgcaaa
     3601 cctggatatc cagaatgtga accaacaaac tgccctacac cttgctgttg aacgacagca
     3661 tacccagatt gttaggcttt tggtccgtgc aggtgccaag cttgatattc aggataagga
     3721 tggggatact cctttgcatg aagctctaag gcatcacact ttgtctcagc tacgtcagct
     3781 ccaagatatg caagatgtgg ggaaggtgga tgctgcctgg gagccatcca aaaacacgtt
     3841 aataatggga cttggtaccc agggggcaga gaagaagagt gcagcatcta ttgcctgttt
     3901 cttggcagcc aatggtgctg acctgagcat tcgaaataag aagggtcaat cgccacttga
     3961 tctctgtcct gatccgaatc tctgcaaagc actggcaaag tgtcataagg aaaaagtcag
     4021 tggtcaagtg ggttctcgga gtccttctat gattagtaat gattctgaaa ccttagaaga
     4081 gtgtatggtg tgctcagata tgaagagaga tactcttttt ggtccatgtg gacatattgc
     4141 tacctgttct ttatgttctc cacgtgtcaa gaaatgcctc atctgtaaag aacaggttca
     4201 atccaggaca aagattgaag aatgtgtggt atgctctgac aagaaagcag ctgttctttt
     4261 tcaaccctgt ggccacatgt gtgcttgtga gaactgtgct aacctgatga aaaagtgtgt
     4321 gcagtgtcga gcagtagttg aacgaagagt gcctttcatt atgtgctgtg gagggaaaag
     4381 ttcagaagat gccactgatg atatctcaag tgggaatatt ccagtattac aaaaggacaa
     4441 ggataatacc aatgtcaatg cagatgtgca aaagttgcag caacagttac aagacattaa
     4501 agagcagaca atgtgccctg tgtgtctaga tcgtctgaag aatatgattt tcctttgtgg
     4561 tcacggaacc tgtcaactct gtggagaccg catgagtgaa tgtcctatct gtcgcaaggc
     4621 tattgaacga aggattcttt tgtatcacca tcaccatcac cattaggatc ctctagaggg
     4681 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     4741 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     4801 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     4861 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     4921 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     4981 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     5041 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     5101 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     5161 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     5221 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5281 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5341 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5401 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     5461 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     5521 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     5581 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5641 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     5701 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     5761 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     5821 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     5881 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     5941 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     6001 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     6061 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     6121 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     6181 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     6241 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     6301 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     6361 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     6421 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     6481 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     6541 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     6601 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     6661 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     6721 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     6781 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6841 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6901 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     6961 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     7021 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     7081 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     7141 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     7201 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     7261 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     7321 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     7381 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7441 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7501 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7561 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7621 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7681 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7741 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     7801 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7861 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac
     7921 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7981 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     8041 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     8101 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     8161 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     8221 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     8281 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     8341 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     8401 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     8461 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8521 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8581 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8641 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8701 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8761 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8821 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8881 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8941 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     9001 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     9061 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     9121 agatctcccg atcccctatg gt
//
