LOCUS       V020424                 7145 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020424
VERSION     V020424
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..1825
                     /label="GNAL(NM_001142339)"
                     /note="GNAL(NM_001142339)"
                     /gene="GNAL"
     CDS             1832..1897
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1921..2509
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2575..2623
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2825..3253
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3267..3596
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3663..4457
                     /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    4633..4754
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4803..4819)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4827..4843
                     /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(4851..4881)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4896..4917
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5205..5793)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5964..6824)
                     /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(6825..6929)
                     /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 ccatggggtg tttgggcggc aacagcaaga cgacggaaga
      721 ccagggcgtc gatgaaaaag aacgacgcga ggccaacaaa aagatcgaga agcagttgca
      781 gaaagagcgc ctggcttaca aggctaccca ccgcctgctg ctcctggggg ctggtgagtc
      841 tgggaaaagc actatcgtca aacagatgag gatcctgcac gtcaatgggt ttaatcccga
      901 ggaaaagaaa cagaaaattc tggacatccg gaaaaatgtt aaagatgcta tcgtgacaat
      961 tgtttcagca atgagtacta taatacctcc agttccgctg gccaaccctg aaaaccaatt
     1021 tcgatcagac tacatcaaga gcatagcccc tatcactgac tttgaatatt cccaggaatt
     1081 ctttgaccat gtgaaaaaac tttgggacga tgaaggcgtg aaggcatgct ttgagagatc
     1141 caacgaatac cagctgattg actgtgcaca atacttcctg gaaagaatcg acagcgtcag
     1201 cttggttgac tacacaccca cagaccagga cctcctcaga tgcagagttc tgacatctgg
     1261 gatttttgag acacgattcc aagtggacaa agtaaacttc cacatgtttg atgttggtgg
     1321 ccagagggat gagaggagaa aatggatcca gtgctttaac gatgtcacag ctatcattta
     1381 cgtcgcagcc tgcagtagct acaacatggt gattcgagaa gataacaaca ccaacaggct
     1441 gagagagtcc ctggatcttt ttgaaagcat ctggaacaac aggtggttac ggaccatttc
     1501 tatcatcttg ttcttgaaca aacaagatat gctggcagaa aaagtcttgg cagggaaatc
     1561 aaaaattgaa gactatttcc cagaatatgc aaattatact gttcctgaag acgcaacacc
     1621 agatgcagga gaagatccca aagttacaag agccaagttc tttatccggg acctgttttt
     1681 gaggatcagc acggccaccg gtgacggcaa acattactgc tacccgcact tcacctgcgc
     1741 cgtggacaca gagaacatcc gcagggtgtt caacgactgc cgcgacatca tccagcggat
     1801 gcacctcaag cagtatgagc tcttgctcga ggactacaaa gaccatgacg gtgattataa
     1861 agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg atatctcgac
     1921 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     1981 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     2041 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     2101 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     2161 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     2221 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     2281 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     2341 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     2401 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     2461 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg
     2521 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     2581 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg
     2641 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     2701 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     2761 aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat
     2821 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2881 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2941 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga
     3001 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3061 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3121 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3181 ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3241 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3301 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3361 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3421 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3481 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     3541 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     3601 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     3661 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3721 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     3781 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     3841 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     3901 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     3961 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4021 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4081 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4141 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     4201 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4261 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4321 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4381 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4441 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc
     4501 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4561 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4621 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4681 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4741 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     4801 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4861 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4921 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4981 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5041 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5101 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5161 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5221 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5281 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5341 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
     5401 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5461 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5521 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5581 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5641 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5701 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5761 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5821 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5881 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5941 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6001 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6061 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6121 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6181 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6241 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6301 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6361 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6421 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6481 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6541 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6601 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6661 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6721 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6781 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6841 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6901 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     6961 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
     7021 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     7081 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     7141 ttagg
//
