LOCUS       V034165                 6995 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034165
VERSION     V034165
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..2488
                     /label="CAT(NM_001752)"
                     /note="CAT(NM_001752)"
                     /gene="CAT"
     CDS             2495..2521
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    2562..2786
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2832..3260
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3274..3603
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3670..4464
                     /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    4638..4759
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4808..4824)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4832..4848
                     /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(4856..4886)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4901..4922
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5210..5795)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5966..6826)
                     /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(6827..6931)
                     /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 gctgacagcc gggatcccgc cagcgaccag atgcagcact ggaaggagca
      961 gcgggccgcg cagaaagctg atgtcctgac cactggagct ggtaacccag taggagacaa
     1021 acttaatgtt attacagtag ggccccgtgg gccccttctt gttcaggatg tggttttcac
     1081 tgatgaaatg gctcattttg accgagagag aattcctgag agagttgtgc atgctaaagg
     1141 agcaggggcc tttggctact ttgaggtcac acatgacatt accaaatact ccaaggcaaa
     1201 ggtatttgag catattggaa agaagactcc catcgcagtt cggttctcca ctgttgctgg
     1261 agaatcgggt tcagctgaca cagttcggga ccctcgtggg tttgcagtga aattttacac
     1321 agaagatggt aactgggatc tcgttggaaa taacaccccc attttcttca tcagggatcc
     1381 catattgttt ccatctttta tccacagcca aaagagaaat cctcagacac atctgaagga
     1441 tccggacatg gtctgggact tctggagcct acgtcctgag tctctgcatc aggtttcttt
     1501 cttgttcagt gatcggggga ttccagatgg acatcgccac atgaatggat atggatcaca
     1561 tactttcaag ctggttaatg caaatgggga ggcagtttat tgcaaattcc attataagac
     1621 tgaccagggc atcaaaaacc tttctgttga agatgcggcg agactttccc aggaagatcc
     1681 tgactatggc atccgggatc tttttaacgc cattgccaca ggaaagtacc cctcctggac
     1741 tttttacatc caggtcatga catttaatca ggcagaaact tttccattta atccattcga
     1801 tctcaccaag gtttggcctc acaaggacta ccctctcatc ccagttggta aactggtctt
     1861 aaaccggaat ccagttaatt actttgctga ggttgaacag atagccttcg acccaagcaa
     1921 catgccacct ggcattgagg ccagtcctga caaaatgctt cagggccgcc tttttgccta
     1981 tcctgacact caccgccatc gcctgggacc caattatctt catatacctg tgaactgtcc
     2041 ctaccgtgct cgagtggcca actaccagcg tgacggcccg atgtgcatgc aggacaatca
     2101 gggtggtgct ccaaattact accccaacag ctttggtgct ccggaacaac agccttctgc
     2161 cctggagcac agcatccaat attctggaga agtgcggaga ttcaacactg ccaatgatga
     2221 taacgttact caggtgcggg cattctatgt gaacgtgctg aatgaggaac agaggaaacg
     2281 tctgtgtgag aacattgccg gccacctgaa ggatgcacaa attttcatcc agaagaaagc
     2341 ggtcaagaac ttcactgagg tccaccctga ctacgggagc cacatccagg ctcttctgga
     2401 caagtacaat gctgagaagc ctaagaatgc gattcacacc tttgtgcagt ccggatctca
     2461 cttggcggca agggagaagg caaatctgct cgagtaccca tacgacgtac cagattacgc
     2521 ttagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     2581 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2641 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     2701 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     2761 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     2821 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2881 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     2941 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3001 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3061 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3121 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3181 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3241 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3301 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3361 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3421 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3481 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3541 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3601 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     3661 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3721 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3781 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3841 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3901 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3961 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4021 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4081 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4141 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4201 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4261 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4321 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4381 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4441 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4501 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4561 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4621 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4681 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4741 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     4801 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4861 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4921 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4981 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5041 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5101 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5161 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5221 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5281 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5341 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5401 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5461 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5521 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5581 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5641 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5701 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca
     5761 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5821 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5881 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5941 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6001 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6061 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6121 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6181 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6241 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6301 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6361 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6421 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6481 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6541 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6601 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6661 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6721 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6781 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6841 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6901 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     6961 tcgacggatc gggagatctc ccgatcccct atggt
//
