LOCUS       V028103                 5768 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028103
VERSION     V028103
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     rep_origin      complement(62..650)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    979..1026
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(1258..2052)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     promoter        complement(2087..2444)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(2446..2550)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    2579..2700
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     CDS             complement(2823..3542)
                     /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"
     promoter        complement(3610..3813)
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     enhancer        3814..4117
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        4118..4321
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     regulatory      4389..4398
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4398..4487
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     CDS             4494..5636
                     /label="KCMF1(NM_020122)"
                     /note="KCMF1(NM_020122)"
                     /gene="KCMF1"
     polyA_signal    5720..5768
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
ORIGIN
        1 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
       61 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      121 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      181 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      241 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      301 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      361 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      421 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      481 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
      541 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      601 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      661 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      721 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      781 aatcaatcta aagtatatat gagtaacctg aggctatggc agggcctgcc gccccgacgt
      841 tggctgcgag ccctgggcct tcacccgaac ttggggggtg gggtggggaa aaggaagaaa
      901 cgcgggcgta ttggccccaa tggggtctcg gtggggtatc gacagagtgc cagccctggg
      961 accgaacccc gcgtttatga acaaacgacc caacaccgtg cgttttattc tgtcttttta
     1021 ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca gttagcctcc
     1081 ccctagggtg ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg
     1141 cgtcccggaa aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat
     1201 ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca
     1261 gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc
     1321 gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt
     1381 agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc
     1441 agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac
     1501 gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag
     1561 cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg
     1621 tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt
     1681 atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga
     1741 tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt
     1801 gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc
     1861 tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg
     1921 gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc
     1981 ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc
     2041 ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcgatctttg caaaagccta
     2101 ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg
     2161 cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg
     2221 gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat
     2281 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg
     2341 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     2401 accctaactg acacacattc cacagctggt tctttccgcc tcaggactct tcctttttca
     2461 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     2521 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgctta
     2581 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     2641 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     2701 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     2761 taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatctaga gtcgcggccg
     2821 ctttacttgt acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc
     2881 aggaccatgt gatcgcgctt ctcgttgggg tctttgctca gggcggactg ggtgctcagg
     2941 tagtggttgt cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg
     3001 tcggcgagct gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg
     3061 ccgttcttct gcttgtcggc catgatatag acgttgtggc tgttgtagtt gtactccagc
     3121 ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc
     3181 accagggtgt cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg
     3241 aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc
     3301 tgcttcatgt ggtcggggta gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg
     3361 agggtgggcc agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg
     3421 ccgtaggtgg catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg
     3481 ccgtccagct cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc
     3541 atggtggcga ccggtggatc ccgggcccgc ggtaccgtcg actgcaggga tctgacggtt
     3601 cactaaacca gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt
     3661 caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa
     3721 actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc
     3781 acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta
     3841 gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg
     3901 ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac
     3961 tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat
     4021 tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg
     4081 tcagccaggc gggccattta ccgtaagtta tgtaacggtg atgcggtttt ggcagtacat
     4141 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     4201 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     4261 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     4321 tctctggcta actagagaac ccactgctta ctggcttatc ggctagcgct accggactca
     4381 gatctcgagc caccatggaa caaaaactaa tatcagagga agatttggaa cagaaactaa
     4441 tcagtgaaga agatttagag caaaagttaa tttccgagga ggacttaact agtatgtccc
     4501 gacatgaagg tgtcagctgt gatgcatgtt taaaaggaaa ttttcgaggt cgcagatata
     4561 agtgtttaat ttgctacgat tacgatcttt gtgcatcttg ttatgaaagt ggtgcaacaa
     4621 caacaaggca tacaactgac cacccaatgc agtgcatatt aacaagggta gattttgatt
     4681 tatactatgg tggggaagct ttctctgtag agcagccaca gtcttttact tgtccctatt
     4741 gtggaaaaat gggctatacg gagacatctc ttcaagaaca tgttacttct gaacatgcag
     4801 aaacatcaac agaagtgatt tgtccaatat gtgcagcgtt acctggaggc gatcctaatc
     4861 atgtcacgga tgactttgca gctcatctta cacttgaaca cagagcccct agagatttag
     4921 atgaatcgag tggtgttcga catgtacgta gaatgtttca ccctggccgg ggattaggag
     4981 gtcctcgtgc tcgtagatca aacatgcact ttactagcag ttctactggt ggactttctt
     5041 cttctcagag ttcatattct ccaagcaata gggaagccat ggatcctata gctgagcttt
     5101 tatctcagtt atcaggagtg agacgttctg caggaggaca gcttaattcc tctggccctt
     5161 ccgcttctca gttacaacaa ctgcagatgc agctgcagct agaacggcag catgcccagg
     5221 cagcacggca acaactggag accgcacgca acgcaacccg gcgtactaac acaagcagtg
     5281 tcaccactac aatcacacaa tccacagcaa caaccaacat agctaataca gaaagcagtc
     5341 agcagactct acagaattcc cagtttcttt taacaaggtt gaatgatcct aaaatgtctg
     5401 aaacggagcg ccagtccatg gaaagcgagc gtgcagaccg cagcctgttt gtccaagagc
     5461 tccttctgtc cactttagtg cgtgaagaga gctcatcctc agatgaggat gatcgggggg
     5521 agatggcaga ttttggtgct atgggctgtg tagatattat gcctttagat gttgctttag
     5581 aaaacctaaa tttaaaagag agtaataaag gaaatgagcc tccaccacct cctctttgag
     5641 aattctcatc tagactggaa atgggggagg ctaactgaaa cacggaagga gacaataccg
     5701 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc
     5761 gtttgttc
//
