LOCUS       V038674                 9993 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038674
VERSION     V038674
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..4673
                     /label="ACE(NM_000789)"
                     /note="ACE(NM_000789)"
                     /gene="ACE"
     CDS             4680..4745
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4769..5357
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5423..5471
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5673..6101
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6115..6444
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6511..7305
                     /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    7481..7602
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7651..7667)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7675..7691
                     /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(7699..7729)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7744..7765
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8053..8641)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8812..9672)
                     /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(9673..9777)
                     /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 ccaccatggg ggccgcctcg ggccgccggg
      781 ggccggggct gctgctgccg ctgccgctgc tgttgctgct gccgccgcag cccgccctgg
      841 cgttggaccc cgggctgcag cccggcaact tttctgctga cgaggccggg gcgcagctct
      901 tcgcgcagag ctacaactcc agcgccgaac aggtgctgtt ccagagcgtg gccgccagct
      961 gggcgcacga caccaacatc accgcggaga atgcaaggcg ccaggaggaa gcagccctgc
     1021 tcagccagga gtttgcggag gcctggggcc agaaggccaa ggagctgtat gaaccgatct
     1081 ggcagaactt cacggacccg cagctgcgca ggatcatcgg agctgtgcgc accctgggct
     1141 ctgccaacct gcccctggct aagcggcagc agtacaacgc cctgctaagc aacatgagca
     1201 ggatctactc caccgccaag gtctgcctcc ccaacaagac tgccacctgc tggtccctgg
     1261 acccagatct caccaacatc ctggcttcct cgcgaagcta cgccatgctc ctgtttgcct
     1321 gggagggctg gcacaacgct gcgggcatcc cgctgaaacc gctgtacgag gatttcactg
     1381 ccctcagcaa tgaagcctac aagcaggacg gcttcacaga cacgggggcc tactggcgct
     1441 cctggtacaa ctcccccacc ttcgaggacg atctggaaca cctctaccaa cagctagagc
     1501 ccctctacct gaacctccat gccttcgtcc gccgcgcact gcatcgccga tacggagaca
     1561 gatacatcaa cctcagggga cccatccctg ctcatctgct gggagacatg tgggcccaga
     1621 gctgggaaaa catctacgac atggtggtgc ctttcccaga caagcccaac ctcgatgtca
     1681 ccagtactat gctgcagcag ggctggaacg ccacgcacat gttccgggtg gcagaggagt
     1741 tcttcacctc cctggagctc tcccccatgc ctcccgagtt ctgggaaggg tcgatgctgg
     1801 agaagccggc cgacgggcgg gaagtggtgt gccacgcctc ggcttgggac ttctacaaca
     1861 ggaaagactt caggatcaag cagtgcacac gggtcacgat ggaccagctc tccacagtgc
     1921 accatgagat gggccatata cagtactacc tgcagtacaa ggatctgccc gtctccctgc
     1981 gtcggggggc caaccccggc ttccatgagg ccattgggga cgtgctggcg ctctcggtct
     2041 ccactcctga acatctgcac aaaatcggcc tgctggaccg tgtcaccaat gacacggaaa
     2101 gtgacatcaa ttacttgcta aaaatggcac tggaaaaaat tgccttcctg ccctttggct
     2161 acttggtgga ccagtggcgc tggggggtct ttagtgggcg taccccccct tcccgctaca
     2221 acttcgactg gtggtatctt cgaaccaagt atcaggggat ctgtcctcct gttacccgaa
     2281 acgaaaccca ctttgatgct ggagctaagt ttcatgttcc aaatgtgaca ccatacatca
     2341 ggtactttgt gagttttgtc ctgcagttcc agttccatga agccctgtgc aaggaggcag
     2401 gctatgaggg cccactgcac cagtgtgaca tctaccggtc caccaaggca ggggccaagc
     2461 tccggaaggt gctgcaggct ggctcctcca ggccctggca ggaggtgctg aaggacatgg
     2521 tcggcttaga tgccctggat gcccagccgc tgctcaagta cttccagcca gtcacccagt
     2581 ggctgcagga gcagaaccag cagaacggcg aggtcctggg ctggcccgag taccagtggc
     2641 acccgccgtt gcctgacaac tacccggagg gcatagacct ggtgactgat gaggctgagg
     2701 ccagcaagtt tgtggaggaa tatgaccgga catcccaggt ggtgtggaac gagtatgccg
     2761 aggccaactg gaactacaac accaacatca ccacagagac cagcaagatt ctgctgcaga
     2821 agaacatgca aatagccaac cacaccctga agtacggcac ccaggccagg aagtttgatg
     2881 tgaaccagtt gcagaacacc actatcaagc ggatcataaa gaaggttcag gacctagaac
     2941 gggcagcact gcctgcccag gagctggagg agtacaacaa gatcctgttg gatatggaaa
     3001 ccacctacag cgtggccact gtgtgccacc cgaatggcag ctgcctgcag ctcgagccag
     3061 atctgacgaa tgtgatggcc acgtcccgga aatatgaaga cctgttatgg gcatgggagg
     3121 gctggcgaga caaggcgggg agagccatcc tccagtttta cccgaaatac gtggaactca
     3181 tcaaccaggc tgcccggctc aatggctatg tagatgcagg ggactcgtgg aggtctatgt
     3241 acgagacacc atccctggag caagacctgg agcggctctt ccaggagctg cagccactct
     3301 acctcaacct gcatgcctac gtgcgccggg ccctgcaccg tcactacggg gcccagcaca
     3361 tcaacctgga ggggcccatt cctgctcacc tgctggggaa catgtgggcg cagacctggt
     3421 ccaacatcta tgacttggtg gtgcccttcc cttcagcccc ctcgatggac accacagagg
     3481 ctatgctaaa gcagggctgg acgcccagga ggatgtttaa ggaggctgat gatttcttca
     3541 cctccctggg gctgctgccc gtgcctcctg agttctggaa caagtcgatg ctggagaagc
     3601 caaccgacgg gcgggaggtg gtctgccacg cctcggcctg ggacttctac aacggcaagg
     3661 acttccggat caagcagtgc accaccgtga acttggagga cctggtggtg gcccaccacg
     3721 aaatgggcca catccagtat ttcatgcagt acaaagactt acctgtggcc ttgagggagg
     3781 gtgccaaccc cggcttccat gaggccattg gggacgtgct agccctctca gtgtctacgc
     3841 ccaagcacct gcacagtctc aacctgctga gcagtgaggg tggcagcgac gagcatgaca
     3901 tcaactttct gatgaagatg gcccttgaca agatcgcctt tatccccttc agctacctcg
     3961 tcgatcagtg gcgctggagg gtatttgatg gaagcatcac caaggagaac tataaccagg
     4021 agtggtggag cctcaggctg aagtaccagg gcctctgccc cccagtgccc aggactcaag
     4081 gtgactttga cccaggggcc aagttccaca ttccttctag cgtgccttac atcaggtact
     4141 ttgtcagctt catcatccag ttccagttcc acgaggcact gtgccaggca gctggccaca
     4201 cgggccccct gcacaagtgt gacatctacc agtccaagga ggccgggcag cgcctggcga
     4261 ccgccatgaa gctgggcttc agtaggccgt ggccggaagc catgcagctg atcacgggcc
     4321 agcccaacat gagcgcctcg gccatgttga gctacttcaa gccgctgctg gactggctcc
     4381 gcacggagaa cgagctgcat ggggagaagc tgggctggcc gcagtacaac tggacgccga
     4441 actccgctcg ctcagaaggg cccctcccag acagcggccg cgtcagcttc ctgggcctgg
     4501 acctggatgc gcagcaggcc cgcgtgggcc agtggctgct gctcttcctg ggcatcgccc
     4561 tgctggtagc caccctgggc ctcagccagc ggctcttcag catccgccac cgcagcctcc
     4621 accggcactc ccacgggccc cagttcggct ccgaggtgga gctgagacac tccggatccg
     4681 actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg
     4741 acaagtagtg agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag
     4801 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     4861 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     4921 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     4981 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     5041 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     5101 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     5161 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     5221 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     5281 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     5341 ttgggccgcc tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata
     5401 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg
     5461 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     5521 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     5581 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga
     5641 tctgcgcagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     5701 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     5761 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     5821 aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     5881 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     5941 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     6001 caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg
     6061 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     6121 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     6181 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     6241 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     6301 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     6361 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     6421 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     6481 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     6541 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     6601 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     6661 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     6721 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     6781 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     6841 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     6901 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     6961 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     7021 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     7081 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     7141 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     7201 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     7261 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     7321 tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     7381 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     7441 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     7501 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     7561 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     7621 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     7681 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     7741 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     7801 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     7861 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     7921 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     7981 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     8041 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     8101 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     8161 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     8221 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     8281 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     8341 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     8401 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     8461 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     8521 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     8581 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     8641 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     8701 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     8761 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     8821 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     8881 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     8941 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     9001 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     9061 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     9121 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     9181 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     9241 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     9301 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     9361 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     9421 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     9481 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     9541 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     9601 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     9661 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     9721 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     9781 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     9841 tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     9901 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     9961 ttgaccgaca attgcatgaa gaatctgctt agg
//
