LOCUS       V046423                 6508 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046423
VERSION     V046423
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             950..2011
                     /label="Dcn(NM_001190451)"
                     /note="Dcn(NM_001190451)"
                     /gene="Dcn"
     CDS             2018..2041
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     promoter        complement(2060..2078)
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /label="SP6 promoter"
     polyA_signal    2104..2328
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2374..2802
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2816..3146
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3213..4007
                     /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    4181..4302
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4351..4367)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4375..4391
                     /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(4399..4429)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4444..4465
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4753..5341)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5512..6372)
                     /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(6373..6477)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcgccacca tgaaggcaac
      961 tctcatcttc ttccttctgg cacaagtctc ttgggctgga ccatttgaac agagaggctt
     1021 atttgacttc atgctagaag atgaggcttc tggcataatc ccttatgacc ctgacaatcc
     1081 cctgatatct atgtgcccct accgatgcca gtgtcatctt cgagtggtgc agtgttctga
     1141 tctgggtttg gacaaagtgc cctgggattt tccacccgac acaaccttgc tagacctgca
     1201 aaacaacaaa attacagaga tcaaagaagg ggccttcaag aacctgaagg acttgcatac
     1261 cttgatcctt gtcaacaaca agatcagcaa aatcagtcca gaggcattca aacctctcgt
     1321 gaagttggaa aggctttacc tgtctaagaa ccaactaaag gaactgcctg aaaaaatgcc
     1381 cagaactctc caggaacttc gtgtccatga gaatgagatc accaagctgc ggaaatccga
     1441 cttcaatgga ctgaacaatg tgcttgtcat agaactgggc ggcaacccac tgaaaaactc
     1501 tgggattgaa aacggagcct tccagggact gaagagtctc tcatacattc gcatctcaga
     1561 caccaacata actgcgatcc ctcaaggtct gcctacttct ctcactgaag tgcatctaga
     1621 tggcaacaag atcaccaagg ttgatgcacc cagcctgaaa ggactgatta atttgtctaa
     1681 actgggattg agcttcaaca gcatcaccgt tatggagaat ggcagtctgg ccaatgttcc
     1741 tcatctgagg gaactccact tggacaacaa caaactcctc agggtgcctg ctgggctggc
     1801 acagcataag tatatccagg tcgtctacct tcacaacaac aacatctccg cagttgggca
     1861 aaatgacttc tgccgagctg gacacccctc tcgaaaggct tcctactcgg ctgtgagtct
     1921 ttacggcaac cctgtccggt attgggaaat ctttccaaac accttcagat gtgtctatgt
     1981 gcgttctgcc attcaacttg gaaactacaa gctcgaggac tacaaggacg acgatgataa
     2041 gtagtctaga gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct
     2101 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     2161 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     2221 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     2281 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     2341 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     2401 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2461 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2521 taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2581 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2641 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     2701 tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt
     2761 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     2821 tcagttaggg tgtggaaagt ccccaggctc cccaggcagg cagaagtatg caaagcatgc
     2881 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     2941 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3001 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3061 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     3121 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     3181 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     3241 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     3301 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     3361 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     3421 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     3481 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     3541 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     3601 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     3661 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     3721 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     3781 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     3841 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     3901 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     3961 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     4021 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4081 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4141 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     4201 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4261 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     4321 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4381 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4441 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4501 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4561 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4621 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4681 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4741 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4801 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4861 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4921 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     4981 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5041 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5101 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5161 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     5221 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5281 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5341 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5401 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5461 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5521 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5581 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5641 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5701 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5761 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5821 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5881 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5941 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6001 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6061 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6121 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6181 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6241 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6301 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6361 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6421 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6481 atttccccga aaagtgccac ctgacgtc
//
