LOCUS       V042996                 7570 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042996
VERSION     V042996
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..3060
                     /label="GFPT1(NM_001244710)"
                     /note="GFPT1(NM_001244710)"
                     /gene="GFPT1"
     CDS             3067..3132
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3170..3394
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3440..3868
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3882..4211
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4278..5072
                     /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    5246..5367
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5416..5432)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5440..5456
                     /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(5464..5494)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5509..5530
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5818..6403)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6574..7434)
                     /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(7435..7539)
                     /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 accatgtgtg gtatatttgc ttacttaaac taccatgttc ctcgaacgag acgagaaatc
     1021 ctggagaccc taatcaaagg ccttcagaga ctggagtaca gaggatatga ttctgctggt
     1081 gtgggatttg atggaggcaa tgataaagat tgggaagcca atgcctgcaa aatccagctt
     1141 attaagaaga aaggaaaagt taaggcactg gatgaagaag ttcacaagca acaagatatg
     1201 gatttggata tagaatttga tgtacacctt ggaatagctc atacccgttg ggcaacacat
     1261 ggagaaccca gtcctgtcaa tagccacccc cagcgctctg ataaaaataa tgaatttatc
     1321 gttattcaca atggaatcat caccaactac aaagacttga aaaagttttt ggaaagcaaa
     1381 ggctatgact tcgaatctga aacagacaca gagacaattg ccaagctcgt taagtatatg
     1441 tatgacaatc gggaaagtca agataccagc tttactacct tggtggagag agttatccaa
     1501 caattggaag gtgcttttgc acttgtgttt aaaagtgttc attttcccgg gcaagcagtt
     1561 ggcacaaggc gaggtagccc tctgttgatt ggtgtacgga gtgaacataa actttctact
     1621 gatcacattc ctatactcta cagaacagct aggactcaga ttggatcaaa attcacacgg
     1681 tggggatcac agggagaaag aggcaaagac aagaaaggaa gctgcaatct ctctcgtgtg
     1741 gacagcacaa cctgcctttt cccggtggaa gaaaaagcag tggagtatta ctttgcttct
     1801 gatgcaagtg ctgtcataga acacaccaat cgcgtcatct ttctggaaga tgatgatgtt
     1861 gcagcagtag tggatggacg tctttctatc catcgaatta aacgaactgc aggagatcac
     1921 cccggacgag ctgtgcaaac actccagatg gaactccagc agatcatgaa gggcaacttc
     1981 agttcattta tgcagaagga aatatttgag cagccagagt ctgtcgtgaa cacaatgaga
     2041 ggaagagtca actttgatga ctatactgtg aatttgggtg gtttgaagga tcacataaag
     2101 gagatccaga gatgccggcg tttgattctt attgcttgtg gaacaagtta ccatgctggt
     2161 gtagcaacac gtcaagttct tgaggagctg actgagttgc ctgtgatggt ggaactagca
     2221 agtgacttcc tggacagaaa cacaccagtc tttcgagatg atgtttgctt tttccttagt
     2281 caatcaggtg agacagcaga tactttgatg ggtcttcgtt actgtaagga gagaggagct
     2341 ttaactgtgg ggatcacaaa cacagttggc agttccatat cacgggagac agattgtgga
     2401 gttcatatta atgctggtcc tgagattggt gtggccagta caaaggctta taccagccag
     2461 tttgtatccc ttgtgatgtt tgcccttatg atgtgtgatg atcggatctc catgcaagaa
     2521 agacgcaaag agatcatgct tggattgaaa cggctgcctg atttgattaa ggaagtactg
     2581 agcatggatg acgaaattca gaaactagca acagaacttt atcatcagaa gtcagttctg
     2641 ataatgggac gaggctatca ttatgctact tgtcttgaag gggcactgaa aatcaaagaa
     2701 attacttata tgcactctga aggcatcctt gctggtgaat tgaaacatgg ccctctggct
     2761 ttggtggata aattgatgcc tgtgatcatg atcatcatga gagatcacac ttatgccaag
     2821 tgtcagaatg ctcttcagca agtggttgct cggcaggggc ggcctgtggt aatttgtgat
     2881 aaggaggata ctgagaccat taagaacaca aaaagaacga tcaaggtgcc ccactcagtg
     2941 gactgcttgc agggcattct cagcgtgatc cctttacagt tgctggcttt ccaccttgct
     3001 gtgctgagag gctatgatgt tgatttccca cggaatcttg ccaaatctgt gactgtagag
     3061 ctcgaggact acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat
     3121 gacgatgaca agtagtgagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     3181 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3241 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3301 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3361 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     3421 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3481 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3541 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3601 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3661 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3721 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3781 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3841 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     3901 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3961 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4021 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4081 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4141 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4201 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     4261 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4321 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4381 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4441 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4501 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4561 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4621 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4681 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4741 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4801 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4861 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4921 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4981 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     5041 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     5101 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5161 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5221 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     5281 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5341 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5401 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5461 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5521 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5581 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5641 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5701 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5761 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5821 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5881 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5941 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6001 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6061 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6121 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6181 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6241 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6301 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt
     6361 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6421 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6481 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6541 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6601 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6661 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6721 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6781 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6841 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6901 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6961 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7021 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7081 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7141 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7201 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7261 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7321 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7381 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7441 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7501 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7561 acctgacgtc
//
