LOCUS       V023800                 6857 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023800
VERSION     V023800
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..2347
                     /label="CFP(NM_001145252)"
                     /note="CFP(NM_001145252)"
                     /gene="CFP"
     CDS             2354..2419
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2457..2681
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2727..3155
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3169..3498
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3565..4359
                     /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    4533..4654
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4703..4719)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4727..4743
                     /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(4751..4781)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4796..4817
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5105..5690)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5861..6721)
                     /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(6722..6826)
                     /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 atgatcacag agggagcgca
      961 ggcccctcga ttgttgctgc cgccgctgct cctgctgctc accctgccag ccacaggctc
     1021 agaccccgtg ctctgcttca cccagtatga agaatcctcc ggcaagtgca agggcctcct
     1081 ggggggtggt gtcagcgtgg aagactgctg tctcaacact gcctttgcct accagaaacg
     1141 tagtggtggg ctctgtcagc cttgcaggtc cccacgatgg tccctgtggt ccacatgggc
     1201 cccctgttcg gtgacgtgct ctgagggctc ccagctgcgg taccggcgct gtgtgggctg
     1261 gaatgggcag tgctctggaa aggtggcacc tgggaccctg gagtggcagc tccaggcctg
     1321 tgaggaccag cagtgctgtc ctgagatggg cggctggtct ggctgggggc cctgggagcc
     1381 ttgctctgtc acctgctcca aagggacccg gacccgcagg cgagcctgta atcaccctgc
     1441 tcccaagtgt gggggccact gcccaggaca ggcacaggaa tcagaggcct gtgacaccca
     1501 gcaggtctgc cccacacacg gggcctgggc cacctggggc ccctggaccc cctgctcagc
     1561 ctcctgccac ggtggacccc acgaacctaa ggagacacga agccgcaagt gttctgcacc
     1621 tgagccctcc cagaaacctc ctgggaagcc ctgcccgggg ctagcctacg agcagcggag
     1681 gtgcaccggc ctgccaccct gcccagtggc tgggggctgg gggccttggg gccctgtgag
     1741 cccctgccct gtgacctgtg gcctgggcca gaccatggaa caacggacgt gcaatcaccc
     1801 tgtgccccag catgggggcc ccttctgtgc tggcgatgcc acccggaccc acatctgcaa
     1861 cacagctgtg ccctgccctg tggatgggga gtgggactcg tggggggagt ggagcccctg
     1921 tatccgacgg aacatgaagt ccatcagctg tcaagaaatc ccgggccagc agtcacgcgg
     1981 gaggacctgc aggggccgca agtttgacgg acatcgatgt gccgggcaac agcaggatat
     2041 ccggcactgc tacagcatcc agcactgccc cttgaaagga tcatggtcag agtggagtac
     2101 ctgggggctg tgcatgcccc cctgtggacc taatcctacc cgtgcccgcc agcgcctctg
     2161 cacacccttg ctccccaagt acccgcccac cgtttccatg gtcgaaggtc agggcgagaa
     2221 gaacgtgacc ttctggggga gaccgctgcc acggtgtgag gagctacaag ggcagaagct
     2281 ggtggtggag gagaaacgac catgtctaca cgtgcctgct tgcaaagacc ctgaggaaga
     2341 ggaactcctc gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta
     2401 caaggatgac gatgacaagt agtgagggcc cgtttaaacc cgctgatcag cctcgactgt
     2461 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2521 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     2581 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     2641 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     2701 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     2761 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     2821 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     2881 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     2941 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3001 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3061 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3121 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3181 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3241 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3301 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3361 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3421 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3481 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     3541 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     3601 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3661 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3721 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     3781 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3841 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3901 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3961 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4021 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4081 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4141 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4201 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4261 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4321 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4381 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4441 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4501 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     4561 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4621 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     4681 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     4741 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     4801 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4861 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4921 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4981 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5041 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5101 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5161 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5221 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5281 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5341 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5401 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5461 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5521 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5581 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5641 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5701 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5761 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5821 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5881 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5941 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6001 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6061 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6121 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6181 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6241 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6301 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6361 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6421 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6481 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6541 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6601 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6661 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6721 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6781 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6841 aagtgccacc tgacgtc
//
