LOCUS       V028941                 8118 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028941
VERSION     V028941
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..2798
                     /label="Gfpt1(NM_013528)"
                     /note="Gfpt1(NM_013528)"
                     /gene="Gfpt1"
     CDS             2805..2870
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2894..3482
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3548..3596
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3798..4226
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4240..4569
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4636..5430
                     /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    5606..5727
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5776..5792)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5800..5816
                     /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(5824..5854)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5869..5890
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6178..6766)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6937..7797)
                     /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(7798..7902)
                     /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 ccaccatgtg cggtatattt gcttatttaa
      781 attaccatgt tcctcgaaca agacgagaaa tcttggagac actaatcaaa ggccttcaga
      841 gactggaata cagaggatat gattctgctg gtgtgggact tgacggaggc aatgacaaag
      901 actgggaagc caacgcctgc aaaatccagc tcattaagaa gaaaggaaaa gttaaggcac
      961 tggatgaaga agttcacaaa caacaagata tggacttgga tatagaattt gatgtgcatc
     1021 ttggaatagc tcatacccgt tgggcgacac atggagaacc caatcctgtc aatagtcacc
     1081 cccagcgctc tgataaaaat aatgaattca ttgttattca taatggaatc atcaccaact
     1141 acaaagactt gaaaaagttt ctggaaagca aaggctatga ctttgaatct gaaacagaca
     1201 cagaaaccat tgccaagctc gtcaagtaca tgtatgacaa ctgggagagc caggacgtca
     1261 gttttaccac cttggtggag agagttatcc aacaattgga aggcgccttt gctcttgtgt
     1321 ttaaaagtgt ccattttccc gggcaagcag ttggcacaag gcgaggtagc cctctcttga
     1381 ttggtgtgcg gagtgaacat aagctttcta cagatcacat tccgattctg tacagaacag
     1441 gcaaagacaa gaaaggaagc tgcggtcttt cccgtgtgga cagcacgaca tgcctgttcc
     1501 ctgttgagga aaaggcagtt gaatattact ttgcttctga tgcaagtgcc gtgatagagc
     1561 acaccaatcg tgtcatcttt ctggaagatg atgatgttgc agcagtggtg gatggccgtc
     1621 tctctatcca ccgaattaaa cgaactgcag gagaccatcc tggccgagct gtgcaaactc
     1681 tccagatgga gctccagcag atcatgaagg gcaactttag ttcatttatg cagaaggaaa
     1741 tttttgagca gccagaatct gttgtgaaca caatgagagg aagagtcaat tttgatgact
     1801 acactgtgaa tttgggaggt ttgaaagatc acattaagga gatccagcgg tgtcggcggt
     1861 tgattcttat tgcttgtggc acaagttacc acgctggtgt ggcaacccgt caggtcctgg
     1921 aggagctgac cgagctgccc gtgatggtgg agcttgccag tgacttcttg gatagaaaca
     1981 ctccagtctt tcgagatgat gtttgctttt tcattagtca atcaggcgag acagctgaca
     2041 ccctgatggg acttcgttac tgtaaggaga gaggagcctt aactgtgggg atcacaaata
     2101 cagtcggcag ttctatatca agggagacag attgcggggt tcatattaat gctggtcctg
     2161 agattggcgt ggccagtaca aaggcataca ccagccagtt tgtgtccctc gtgatgtttg
     2221 ctctcatgat gtgtgatgac aggatctcca tgcaagagag acgcaaagag atcatgctcg
     2281 gactgaagcg actgccggac ttgattaagg aagtgctgag catggatgat gaaatccaga
     2341 agctggcgac ggagctttac caccagaagt cggtcctgat aatggggcgg ggctaccatt
     2401 atgctacatg ccttgaaggg gctctgaaaa tcaaggagat tacttatatg cattcggaag
     2461 gcatccttgc tggtgagctc aagcacggcc ctctggcctt ggtggacaag ttgatgcctg
     2521 tcatcatgat catcatgcga gaccacactt atgccaagtg ccagaacgct cttcagcagg
     2581 tggttgcacg gcaggggcgt ccagtcgtga tctgtgataa ggaggatact gagaccatta
     2641 agaatacaaa aaggacaatc aaggtgcccc actcagtgga ctgcttgcag ggcattctca
     2701 gtgtgattcc cctgcagctg ctggctttcc acctggctgt gctgagaggc tacgatgttg
     2761 attttccacg gaatcttgcc aaatctgtaa cagtagaggg atccgactac aaagaccatg
     2821 acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc
     2881 ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     2941 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     3001 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     3061 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     3121 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     3181 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     3241 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     3301 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     3361 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     3421 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     3481 gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     3541 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     3601 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     3661 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     3721 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg
     3781 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3841 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3901 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     3961 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4021 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4081 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4141 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     4201 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     4261 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4321 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4381 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4441 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4501 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4561 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     4621 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4681 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4741 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4801 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4861 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4921 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4981 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     5041 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5101 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5161 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5221 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5281 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5341 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5401 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga
     5461 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5521 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5581 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     5641 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5701 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5761 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5821 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5881 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5941 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6001 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6061 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6121 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6181 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6241 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6301 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6361 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6421 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6481 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6541 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6601 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6661 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6721 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6781 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6841 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6901 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6961 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7021 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7081 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7141 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7201 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7261 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7321 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7381 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7441 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7501 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7561 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7621 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7681 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7741 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7801 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7861 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7921 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca
     7981 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     8041 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     8101 atgaagaatc tgcttagg
//
