LOCUS       V023810                 6209 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023810
VERSION     V023810
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..1699
                     /label="CFD(NM_001928)"
                     /note="CFD(NM_001928)"
                     /gene="CFD"
     CDS             1706..1771
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    1809..2033
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2079..2507
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2521..2850
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             2917..3711
                     /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    3885..4006
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4055..4071)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4079..4095
                     /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(4103..4133)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4148..4169
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4457..5042)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5213..6073)
                     /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(6074..6178)
                     /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 atgcacagct gggagcgcct
      961 ggcagttctg gtcctcctag gagcggccgc ctgcgcggcg ccgccccgtg gtcggatcct
     1021 gggcggcaga gaggccgagg cgcacgcgcg gccctacatg gcgtcggtgc agctgaacgg
     1081 cgcgcacctg tgcggcggcg tcctggtggc ggagcagtgg gtgctgagcg cggcgcactg
     1141 cctggaggac gcggccgacg ggaaggtgca ggttctcctg ggcgcgcact ccctgtcgca
     1201 gccggagccc tccaagcgcc tgtacgacgt gctccgcgca gtgccccacc cggacagcca
     1261 gcccgacacc atcgaccacg acctcctgct gctacagctg tcggagaagg ccacactggg
     1321 ccctgctgtg cgccccctgc cctggcagcg cgtggaccgc gacgtggcac cgggaactct
     1381 ctgcgacgtg gccggctggg gcatagtcaa ccacgcgggc cgccgcccgg acagcctgca
     1441 gcacgtgctc ttgccagtgc tggaccgcgc cacctgcaac cggcgcacgc accacgacgg
     1501 cgccatcacc gagcgcttga tgtgcgcgga gagcaatcgc cgggacagct gcaagggtga
     1561 ctccgggggc ccgctggtgt gcgggggcgt gctcgagggc gtggtcacct cgggctcgcg
     1621 cgtttgcggc aaccgcaaga agcccgggat ctacacccgc gtggcgagct atgcggcctg
     1681 gatcgacagc gtcctggccc tcgaggacta caaagaccat gacggtgatt ataaagatca
     1741 tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgtttaaa cccgctgatc
     1801 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     1861 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     1921 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     1981 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     2041 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     2101 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     2161 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2221 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2281 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     2341 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     2401 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     2461 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     2521 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2581 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     2641 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     2701 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     2761 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     2821 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     2881 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     2941 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3001 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3061 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     3121 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3181 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3241 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     3301 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     3361 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     3421 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     3481 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     3541 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     3601 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     3661 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     3721 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     3781 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     3841 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     3901 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3961 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     4021 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4081 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4141 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4201 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4261 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4321 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4381 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4441 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4501 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4561 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4621 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4681 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4741 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4801 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4861 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4921 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4981 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5041 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5101 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5161 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5221 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5281 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5341 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5401 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5461 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5521 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5581 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5641 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5701 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5761 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5821 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5881 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5941 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6001 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6061 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6121 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6181 catttccccg aaaagtgcca cctgacgtc
//
