LOCUS       V027457                 7791 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027457
VERSION     V027457
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..2471
                     /label="FTCD(NM_001320412)"
                     /note="FTCD(NM_001320412)"
                     /gene="FTCD"
     CDS             2478..2543
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2567..3155
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3221..3269
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3471..3899
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3913..4242
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4309..5103
                     /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    5279..5400
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5449..5465)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5473..5489
                     /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(5497..5527)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5542..5563
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5851..6439)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6610..7470)
                     /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(7471..7575)
                     /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 ccaccatgtc ccagctggtg gaatgcgtcc
      781 ccaacttttc ggaggggaag aaccaggagg tgatcgacgc catctctgga gccatcacac
      841 agaccccggg ctgcgtgctg ctggatgtgg acgcaggccc ttccaccaac cgcaccgtgt
      901 acaccttcgt ggggccgccg gagtgcgtgg tggagggggc cctcaacgct gcccgggtag
      961 cttcccgact tatcgacatg agcaggcacc aaggagagca cccccgcatg ggggccctag
     1021 acgtctgccc cttcatcccc gtgaggggcg tcagcgtgga tgagtgtgtg ctctgcgccc
     1081 aggcctttgg ccagaggctg gcagaggagc tggacgtgcc agtttacctg tacggcgagg
     1141 cagccaggat ggacagtcgc cggaccctgc cggccatccg ggccggggag tacgaggccc
     1201 tccctaagaa gctccagcag gccgactggg cgcccgactt tggtcccagc tcctttgtcc
     1261 ccagttgggg ggccacggcc acgggggcga ggaagttcct cattgctttt aacatcaacc
     1321 tgctcggcac aaaggagcaa gcccaccgca tcgcgctcaa cctgcgggag cagggccgcg
     1381 ggaaggacca gccaggacgt ctgaagaaag ttcagggcat tggctggtac ctggatgaga
     1441 agaacctggc tcaggtgtcc accaatcttc tggactttga ggtcacggca ctgcacacgg
     1501 tctacgagga gacctgccga gaagcacagg agctgagcct cccagtggtg ggctcacagc
     1561 tggtgggcct ggtgcccctg aaggctctgc tggatgcggc cgccttctac tgcgagaagg
     1621 agaacctctt catcctggag gaggagcagc ggatcaggct ggtggtgagc cggctgggcc
     1681 tggactccct gtgccccttc agccctaagg agcggatcat cgagtacctg gtccctgagc
     1741 gcgggcctga gcgaggcctg ggcagcaagt ccctgcgcgc cttcgtgggg gaggtgggtg
     1801 cccgctctgc ggcccccggg ggcggctcgg tggcggcggc cgctgcggcc atgggtgcgg
     1861 cgctgggctc catggtgggc ctcatgacct acgggcggcg ccaattccag tccctggaca
     1921 cgacgatgcg gcgcctgatc ccgcccttcc gcgaggcttc ggccaagcta accacgctgg
     1981 tggatgccga cgccgaggcc ttcaccgcct acctggaagc aatgaggctc cccaagaaca
     2041 cacctgagga aaaggacagg cgcacggcgg ccctacagga gggtctgagg cgggcagtct
     2101 ctgtgccgct gacgctggcg gagacggtgg cctcgctgtg gccggccctg caggaactgg
     2161 cccggtgtgg gaacctggcc tgccggtcag acctccaggt ggcggccaaa gccctggaga
     2221 tgggcgtgtt tggcgcatat ttcaacgtgc tcatcaacct gagggacatc acagacgagg
     2281 catttaagga ccagcctcct gcaggaagcc aagacccagg ctgcactggt gctggactgc
     2341 ttggagaccc ggcaggagtg acggtgcggg aggcctctcc gggctctgtg gcccctccct
     2401 cgcccatccc cagagggcag agctgtgacc tcgaaacccc agggacagca ggaccttcga
     2461 cgctggaagg aggatccgac tacaaagacc atgacggtga ttataaagat catgacatcg
     2521 actacaagga tgacgatgac aagtagtgag ggcccgatat ctcgacaatc aacctctgga
     2581 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2641 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2701 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     2761 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     2821 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     2881 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     2941 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     3001 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3061 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3121 gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa
     3181 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     3241 aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     3301 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     3361 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     3421 aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg
     3481 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3541 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3601 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
     3661 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3721 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3781 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3841 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3901 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3961 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4021 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4081 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4141 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     4201 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     4261 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4321 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4381 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4441 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4501 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4561 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4621 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4681 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4741 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4801 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4861 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4921 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4981 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5041 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5101 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     5161 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5221 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     5281 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5341 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5401 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     5461 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5521 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5581 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5641 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5701 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5761 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5821 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5881 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5941 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6001 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     6061 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6121 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6181 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6241 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     6301 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6361 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6421 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6481 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6541 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6601 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6661 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6721 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6781 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6841 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6901 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6961 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7021 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7081 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7141 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7201 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7261 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7321 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7381 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7441 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7501 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7561 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     7621 tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc
     7681 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     7741 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
