LOCUS       V030327                 7019 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030327
VERSION     V030327
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             941..2509
                     /label="GPT2(NM_133443)"
                     /note="GPT2(NM_133443)"
                     /gene="GPT2"
     CDS             2516..2581
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2619..2843
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2889..3317
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3331..3660
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3727..4521
                     /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    4695..4816
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4865..4881)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4889..4905
                     /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(4913..4943)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4958..4979
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5267..5852)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6023..6883)
                     /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(6884..6988)
                     /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 atccgccacc atgcagcggg cggcggcgct
      961 ggtccggcgg ggctgtggtc cccggacccc cagctcctgg ggccgcagcc agagcagcgc
     1021 ggccgccgag gcctcggcgg tgctcaaggt gcggcccgag cgcagccggc gcgagcgcat
     1081 cctcacgctg gagtccatga acccgcaggt gaaggcggtg gagtacgccg tgcggggacc
     1141 catcgtgctc aaggccggcg agatcgagct cgagctgcag cggggtatca aaaagccatt
     1201 cacagaggtc atccgagcca acatcgggga cgcccaggct atggggcagc agccaatcac
     1261 cttcctccgg caggtgatgg cactatgcac ctacccaaac ctgctggaca gccccagctt
     1321 cccagaagat gctaagaaac gtgcccggcg gatcctgcag gcttgtggcg ggaacagcct
     1381 ggggtcctac agtgctagcc agggtgtcaa ctgcatccgt gaagatgtgg ctgcctacat
     1441 caccaggagg gatggcggtg tgcctgcgga ccccgacaac atctacctga ccacgggagc
     1501 tagtgacggc atttctacga tcctgaagat cctcgtctcc gggggcggca agtcacggac
     1561 aggtgtgatg atccccatcc cacaatatcc cctctattca gctgtcatct ctgagctcga
     1621 cgccatccag gtgaattact acctggacga ggagaactgc tgggcgctga atgtgaatga
     1681 gctccggcgg gcggtgcagg aggccaaaga ccactgtgat cctaaggtgc tctgcataat
     1741 caaccctggg aaccccacag gccaggtaca aagcagaaag tgcatagaag atgtgatcca
     1801 ctttgcctgg gaagagaagc tctttctcct ggctgatgag gtgtaccagg acaacgtgta
     1861 ctctccagat tgcagattcc actccttcaa gaaggtgctg tacgagatgg ggcccgagta
     1921 ctccagcaac gtggagctcg cctccttcca ctccacctcc aagggctaca tgggcgagtg
     1981 tggttacaga ggaggctaca tggaggtgat caacctgcac cctgagatca agggccagct
     2041 ggtgaagctg ctgtcggtgc gcctgtgccc cccagtgtct gggcaggccg ccatggacat
     2101 tgtcgtgaac cccccggtgg caggagagga gtcctttgag caattcagcc gagagaagga
     2161 gtcggtcctg ggtaatctgg ccaaaaaagc aaagctgacg gaagacctgt ttaaccaagt
     2221 cccaggaatt cactgcaacc ccttgcaggg ggccatgtac gccttccctc ggatcttcat
     2281 tcctgccaaa gctgtggagg ctgctcaggc ccatcaaatg gctccagaca tgttctactg
     2341 catgaagctc ctggaggaga ctggcatctg tgtcgtgccc ggcagtggct ttgggcagag
     2401 ggaaggcact taccacttca ggatgactat cctccctcca gtggagaagc tgaaaacggt
     2461 gctgcagaag gtgaaagact tccacatcaa cttcctggag aagtacgcgc tcgaggacta
     2521 caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa
     2581 gtagtgaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     2641 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     2701 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     2761 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     2821 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     2881 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2941 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3001 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3061 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3121 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3181 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3241 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3301 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3361 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3421 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3481 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3541 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     3601 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3661 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     3721 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3781 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3841 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3901 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3961 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4021 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4081 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4141 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4201 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4261 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4321 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4381 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4441 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4501 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4561 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4621 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4681 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     4741 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     4801 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     4861 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     4921 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     4981 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5041 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5101 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5161 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5221 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5281 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5341 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5401 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5461 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5521 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5581 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5641 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5701 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5761 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc
     5821 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5881 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5941 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6001 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6061 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6121 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6181 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6241 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6301 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6361 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6421 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6481 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6541 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6601 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6661 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6721 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6781 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6841 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6901 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6961 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
