LOCUS       V034787                 7693 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034787
VERSION     V034787
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             964..3183
                     /label="POLI(NM_007195)"
                     /note="POLI(NM_007195)"
                     /gene="POLI"
     CDS             3190..3255
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3293..3517
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3563..3991
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4005..4334
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4401..5195
                     /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    5369..5490
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5539..5555)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5563..5579
                     /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(5587..5617)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5632..5653
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5941..6526)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6697..7557)
                     /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(7558..7662)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggaga agctgggggt ggagccggag gaggaaggcg gcggcgacga cgacgaggaa
     1021 gacgccgagg cctgggccat ggaactggcg gacgtggggg cggcagccag ctcgcaggga
     1081 gttcatgatc aagtgttgcc cacaccaaat gcttcatcca gagtcatagt acatgtggat
     1141 ctggattgct tttatgcaca agtagaaatg atctcaaatc cagagctaaa agacaaacct
     1201 ttaggggttc aacagaaata tttggtggtt acctgcaact atgaagctag gaaacttgga
     1261 gttaagaaac ttatgaatgt cagagatgca aaagaaaagt gtccacagtt ggtattagtt
     1321 aatggagaag acctgacccg ctacagagaa atgtcttata aggttacaga attactggaa
     1381 gaatttagtc cagttgttga gagacttgga tttgatgaaa attttgtgga tctaacagaa
     1441 atggttgaga agagactaca gcagctgcaa agtgatgaac tttctgcggt gactgtgtcg
     1501 ggtcatgtat acaataatca gtctataaac ctgcttgacg tcttgcacat cagactactt
     1561 gttggatctc agattgcagc agagatgcgg gaagccatgt ataatcagtt ggggctcact
     1621 ggctgtgctg gagtggcttc taataaactg ttggcaaaat tagtttctgg tgtctttaaa
     1681 ccaaatcaac aaacagtctt attacctgaa agttgtcaac atcttattca tagtttgaat
     1741 cacataaagg aaatacctgg tattggctat aaaactgcca aatgtcttga agcactgggt
     1801 atcaatagtg tgcgtgatct ccaaaccttt tcacccaaaa ttttagaaaa agaattagga
     1861 atttcagttg ctcagcgtat ccaaaagctc agttttggag aggataactc ccctgtgata
     1921 ctctcaggac cacctcagtc ctttagtgaa gaagattcat ttaaaaaatg ttcatctgaa
     1981 gttgaagcta aaaataagat tgaagaacta cttgctagtc ttttaaacag agtatgccaa
     2041 gatggaagga agcctcatac agtgagatta ataatccgtc ggtattcctc tgagaagcac
     2101 tatggtcgtg agagtcgtca gtgccctatt ccttcacatg taattcagaa attagggaca
     2161 ggaaattatg atgtgatgac cccaatggtt gatatactta tgaaactttt tcgaaatatg
     2221 gtgaatgtga agatgccatt tcaccttacc cttctaagtg tgtgcttctg caaccttaaa
     2281 gcactaaata ctgctaagaa agggcttatt gattattatt taatgccatc attatcaact
     2341 acttcacgct ctggcaagca cagttttaaa atgaaagaca ctcatatgga agattttccc
     2401 aaagacaaag aaacaaaccg ggatttccta ccaagtggaa gaattgaaag tacaagaact
     2461 agggagtctc cactagatac cacaaatttt tctaaagaaa aagacattaa tgaattccca
     2521 ctctgttcac ttcctgaagg tgttgaccaa gaagtcttca agcagcttcc agtagatatt
     2581 caagaagaaa tcctttctgg aaaatctagg gaaaaatttc aagggaaagg aagtgtgagt
     2641 tgtccattac atgcctctag aggagtatta tctttctttt ctaaaaaaca aatgcaagat
     2701 attcccataa atcctagaga tcatttatcc agtagcaaac aggtatcctc tgtatctcct
     2761 tgtgaaccgg gaacatcagg ctttaatagc agtagttctt cttacatgtc tagccaaaag
     2821 gattattcat attatttaga taatagatta aaagatgaac gaataagtca aggacctaaa
     2881 gaacctcaag gattccactt tacaaattca aaccctgctg tgtctgcttt tcattcattt
     2941 ccaaacttgc agagtgagca acttttctcc agaaaccaca ctacagatag ccataagcaa
     3001 acagtagcaa cagactctca tgaaggactt acagaaaata gagagccaga ttctgttgat
     3061 gagaaaatta ctttcccttc tgacattgat cctcaagttt tctatgaact accagaagca
     3121 gtacaaaagg aactgctggc agagtggaag agagcaggat cagatttcca cattggacat
     3181 aaactcgagg actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag
     3241 gatgacgatg acaagtagtg agggcccgtt taaacccgct gatcagcctc gactgtgcct
     3301 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3361 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3421 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3481 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3541 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3601 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3661 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3721 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3781 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3841 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3901 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3961 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     4021 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4081 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4141 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4201 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4261 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4321 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     4381 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4441 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4501 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4561 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4621 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4681 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4741 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4801 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4861 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4921 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4981 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5041 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5101 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5161 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5221 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5281 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5341 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     5401 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5461 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     5521 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5581 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5641 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5701 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5761 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5821 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5881 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5941 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6001 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6061 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6121 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6181 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6241 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6301 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6361 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6421 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt
     6481 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6541 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6601 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6661 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6721 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6781 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6841 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6901 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6961 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7021 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7081 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7141 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7201 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7261 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7321 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7381 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7441 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7501 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7561 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7621 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7681 gccacctgac gtc
//
