LOCUS       V019795                 7634 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019795
VERSION     V019795
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"
     CDS             683..2314
                     /label="Cdyl(NM_001123386)"
                     /note="Cdyl(NM_001123386)"
                     /gene="Cdyl"
     CDS             2321..2386
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2410..2998
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3064..3112
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3314..3742
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3756..4085
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4152..4946
                     /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    5122..5243
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5292..5308)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5316..5332
                     /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(5340..5370)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5385..5406
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5694..6282)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6453..7313)
                     /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(7314..7418)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatggcctc cgaggagctg tacgaggtgg aaagtatcgt
      721 tgacaaaagg aaaaacaaga aagggaagac agaatatctg gtgcggtgga aaggctatga
      781 cagtgaggat gacacgtggg agcctgagca gcacctggtg aactgtgagg aatacatcca
      841 tgacttcaac cggcgccaca acgagaggca aaaggaaggt agcctggctc gtgccagcag
      901 agcctccccc agcaacgccc ggaagcagat ttccaggtcc acccacagca ctctctccaa
      961 gaccaactcc aaagcacttg tggtaggcaa agatcatgag tccaaaagca gccagctgtt
     1021 ggctgccagc cagaagttca ggaaaaaccc agccccatct cttgcaaacc gcaagaacat
     1081 ggacctcgcc aagtcaggga tcaaaattct cgtgcctaag agccccgtta agggcaggac
     1141 ctcggttgat ggctttcagg gggagagccc cgagaagctg gaccctgtgg atcagggtgc
     1201 cgaggacact gtagccccag aggtgactgc agagaagccc actggggctt tgctgggccc
     1261 tggtgcggag cgagccagga tggggagcag gccccgaata catccactag tgcctcaggt
     1321 ttctggcccc gtgactgctg ccatggccac aggcttagct gttaatggaa aaggtacatc
     1381 tccattcatg gatgcgctag cagccaacgg aacagtcacc atacagacat ccgtaacagg
     1441 agtgacagcc gggaaaagga aatttattga cgacagaaga gaccaacctt ttgacaagcg
     1501 gttgcgtttc agtgtgaggc agacagagag tgcctacaga tacagagata ttgtcgtcag
     1561 gaagcaagat ggcttcaccc acatcttgtt atccacaaaa tcgtcagaga ataactcact
     1621 aaacccagag gtgatgaaag aagtgcagag cgccctgagc acagctgcag ccgacgacag
     1681 caagctggtt ctgctcagcg ccgtgggcag cgtcttctgc tgtggtctgg actttattta
     1741 ttttattcgg cgcctcacag atgaccgaaa gagagaaagc actaaaatgg cagacgctat
     1801 cagaaacttc gtgaatactt tcattcagtt taagaagcct attattgtag ctgttaatgg
     1861 cccagccatt ggactaggag catccatatt gcctctttgt gatgtggttt gggctaacga
     1921 aaaggcttgg tttcaaacac cctataccac cttcggacag agtccagatg gctgctctac
     1981 cgttatgttt cccaagatta tgggaggagc atctgcgaat gaaatgctgt tcagtgggcg
     2041 gaagttgacg gcacaggagg cctgtggcaa gggtctggtc tcccaggtgt tttggccagg
     2101 aaccttcaca caggaagtca tggttcgaat caaggagctg gcttcatgta acccagttgt
     2161 cctggaggaa tccaaagccc tggtgcgctg caatatgaag atggagctag agcaggccaa
     2221 tgagagagaa tgtgaagtgc tgaagaagat ctggggctcc gcccagggca tggactccat
     2281 gttaaagtac ttacagagga aaatcgatga gttcctcgag gactacaaag accatgacgg
     2341 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccga
     2401 tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2461 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2521 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     2581 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     2641 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     2701 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     2761 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     2821 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     2881 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     2941 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     3001 ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     3061 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc
     3121 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     3181 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     3241 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct
     3301 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3361 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3421 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta
     3481 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3541 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3601 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3661 tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt
     3721 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3781 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3841 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3901 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3961 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4021 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4081 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     4141 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4201 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4261 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4321 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4381 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4441 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4501 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4561 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4621 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4681 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4741 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4801 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4861 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4921 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa
     4981 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5041 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5101 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     5161 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5221 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5281 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5341 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5401 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5461 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5521 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5581 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5641 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5701 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5761 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5821 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5881 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5941 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6001 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6061 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6121 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6181 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6241 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6301 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6361 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6421 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6481 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6541 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6601 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6661 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6721 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6781 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6841 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6901 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6961 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7021 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7081 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7141 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7201 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7261 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7321 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7381 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7441 cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct
     7501 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     7561 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     7621 agaatctgct tagg
//
