LOCUS       V021945                 8716 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021945
VERSION     V021945
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             649..1365
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             1399..3426
                     /label="KCNQ1(NM_000218)"
                     /note="KCNQ1(NM_000218)"
                     /gene="KCNQ1"
     misc_feature    3492..4080
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4146..4194
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4396..4824
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4838..5167
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5234..6028
                     /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    6204..6325
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6374..6390)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6398..6414
                     /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(6422..6452)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6467..6488
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6776..7364)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7535..8395)
                     /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(8396..8500)
                     /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 gcgccaccat ggtgagcaag
      661 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      721 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      781 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      841 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
      901 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
      961 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1021 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1081 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     1141 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     1201 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     1261 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1321 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggagg agggggaagc
     1381 ggtaccgagc tcggatccat ggccgcggcc tcctccccgc ccagggccga gaggaagcgc
     1441 tggggttggg gccgcctgcc aggcgcccgg cggggcagcg cgggcctggc caagaagtgc
     1501 cccttctcgc tggagctggc ggagggcggc ccggcgggcg gcgcgctcta cgcgcccatc
     1561 gcgcccggcg ccccaggtcc cgcgccccct gcgtccccgg ccgcgcccgc cgcgccccca
     1621 gttgcctccg accttggccc gcggccgccg gtgagcctag acccgcgcgt ctccatctac
     1681 agcacgcgcc gcccggtgtt ggcgcgcacc cacgtccagg gccgcgtcta caacttcctc
     1741 gagcgtccca ccggctggaa atgcttcgtt taccacttcg ccgtcttcct catcgtcctg
     1801 gtctgcctca tcttcagcgt gctgtccacc atcgagcagt atgccgccct ggccacgggg
     1861 actctcttct ggatggagat cgtgctggtg gtgttcttcg ggacggagta cgtggtccgc
     1921 ctctggtccg ccggctgccg cagcaagtac gtgggcctct gggggcggct gcgctttgcc
     1981 cggaagccca tttccatcat cgacctcatc gtggtcgtgg cctccatggt ggtcctctgc
     2041 gtgggctcca aggggcaggt gtttgccacg tcggccatca ggggcatccg cttcctgcag
     2101 atcctgagga tgctacacgt cgaccgccag ggaggcacct ggaggctcct gggctccgtg
     2161 gtcttcatcc tccgccagga gctgataacc accctgtaca tcggcttcct gggcctcatc
     2221 ttctcctcgt actttgtgta cctggctgag aaggacgcgg tgaacgagtc aggccgcgtg
     2281 gagttcggca gctacgcaga tgcgctgtgg tggggggtgg tcacagtcac caccatcggc
     2341 tatggggaca aggtgcccca gacgtgggtc gggaagacca tcgcctcctg cttctctgtc
     2401 tttgccatct ccttctttgc gctcccagcg gggattcttg gctcggggtt tgccctgaag
     2461 gtgcagcaga agcagaggca gaagcacttc aaccggcaga tcccggcggc agcctcactc
     2521 attcagaccg catggaggtg ctatgctgcc gagaaccccg actcctccac ctggaagatc
     2581 tacatccgga aggccccccg gagccacact ctgctgtcac ccagccccaa acccaagaag
     2641 tctgtggtgg taaagaaaaa aaagttcaag ctggacaaag acaatggggt gactcctgga
     2701 gagaagatgc tcacagtccc ccatatcacg tgcgaccccc cagaagagcg gcggctggac
     2761 cacttctctg tcgacggcta tgacagttct gtaaggaaga gcccaacact gctggaagtg
     2821 agcatgcccc atttcatgag aaccaacagc ttcgccgagg acctggacct ggaaggggag
     2881 actctgctga cacccatcac ccacatctca cagctgcggg aacaccatcg ggccaccatt
     2941 aaggtcattc gacgcatgca gtactttgtg gccaagaaga aattccagca agcgcggaag
     3001 ccttacgatg tgcgggacgt cattgagcag tactcgcagg gccacctcaa cctcatggtg
     3061 cgcatcaagg agctgcagag gaggctggac cagtccattg ggaagccctc actgttcatc
     3121 tccgtctcag aaaagagcaa ggatcgcggc agcaacacga tcggcgcccg cctgaaccga
     3181 gtagaagaca aggtgacgca gctggaccag aggctggcac tcatcaccga catgcttcac
     3241 cagctgctct ccttgcacgg tggcagcacc cccggcagcg gcggcccccc cagagagggc
     3301 ggggcccaca tcacccagcc ctgcggcagt ggcggctccg tcgaccctga gctcttcctg
     3361 cccagcaaca ccctgcccac ctacgagcag ctgaccgtgc ccaggagggg ccccgatgag
     3421 gggtcctgag aattctgcag atatccagca cagtggcggc cgctcgagtc tagagggccc
     3481 gatatctcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     3541 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     3601 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     3661 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     3721 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     3781 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     3841 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     3901 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     3961 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     4021 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     4081 ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     4141 tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac
     4201 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     4261 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     4321 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct
     4381 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4441 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     4501 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt
     4561 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     4621 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4681 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     4741 attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga
     4801 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     4861 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4921 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4981 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     5041 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     5101 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     5161 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     5221 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     5281 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     5341 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     5401 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     5461 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     5521 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     5581 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     5641 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     5701 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     5761 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5821 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5881 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5941 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     6001 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc
     6061 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     6121 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     6181 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     6241 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6301 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     6361 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     6421 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     6481 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6541 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     6601 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     6661 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6721 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6781 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6841 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6901 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6961 cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     7021 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     7081 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     7141 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     7201 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     7261 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     7321 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     7381 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     7441 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     7501 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     7561 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     7621 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     7681 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     7741 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     7801 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     7861 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     7921 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7981 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     8041 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     8101 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     8161 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     8221 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     8281 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     8341 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     8401 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     8461 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     8521 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat
     8581 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     8641 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     8701 gaagaatctg cttagg
//
