LOCUS       V020026                 7881 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020026
VERSION     V020026
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        236..615
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        616..819
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        864..882
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             965..2665
                     /label="LCTL(NM_207338)"
                     /note="LCTL(NM_207338)"
                     /gene="LCTL"
     CDS             2711..3430
                     /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"
     polyA_signal    3482..3706
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3752..4180
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4194..4523
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4590..5384
                     /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    5558..5679
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5728..5744)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5752..5768
                     /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(5776..5806)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5821..5842
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6130..6715)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6886..7746)
                     /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(7747..7851)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
       61 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
      121 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
      181 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
      241 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
      301 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      361 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      421 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      481 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      541 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      601 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      661 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      721 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      781 ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc
      841 actgcttact ggcttatcga aattaatacg actcactata gggagaccca agctggctag
      901 cgtttaaact taagcttggt accgagctcg gatccactag tccagtgtgg tggaattcgc
      961 caccatgaag ccagtgtggg tcgccaccct tctgtggatg ctactgctgg tgcccaggct
     1021 gggggccgcc cggaaggggt ccccagaaga ggcctccttc tactatggaa ccttccctct
     1081 tggcttctcc tggggcgtgg gcagttctgc ctaccagacg gagggcgcct gggaccagga
     1141 cgggaaaggg cctagcatct gggacgtctt cacacacagt gggaagggga aagtgcttgg
     1201 gaatgagacg gcagatgtag cctgtgacgg ctactacaag gtccaggagg acatcattct
     1261 gctgagggaa ctgcacgtca accactaccg attctccctg tcttggcccc ggctcctgcc
     1321 cacaggcatc cgagccgagc aggtgaacaa gaagggaatc gaattctaca gtgatcttat
     1381 cgatgccctt ctgagcagca acatcactcc catcgtgacc ttgcaccact gggatctgcc
     1441 acagctgctc caggtcaaat acggtgggtg gcagaatgtg agcatggcca actacttcag
     1501 agactacgcc aacctgtgct ttgaggcctt tggggaccgt gtgaagcact ggatcacgtt
     1561 cagtgatcct cgggcaatgg cagaaaaagg ctatgagacg ggccaccatg cgccgggcct
     1621 gaagctccgc ggcaccggcc tgtacaaggc agcacaccac atcattaagg cccacgccaa
     1681 agcctggcat tcttataaca ccacgtggcg cagcaagcag caaggtctgg tgggaatttc
     1741 attgaactgt gactgggggg aacctgtgga cattagtaac cccaaggacc tagaggctgc
     1801 cgagagatac ctacagttct gtctgggctg gtttgccaac cccatttatg ccggtgacta
     1861 cccccaagtc atgaaggact acattggaag aaagagtgca gagcaaggcc tggagatgtc
     1921 gaggttaccg gtgttctcac tccaggagaa gagctacatt aaaggcacat ccgatttctt
     1981 gggattaggt cattttacta ctcggtacat cacggaaagg aactacccct cccgccaggg
     2041 gcccagctac cagaacgatc gtgacttgat agagctggtt gacccaaact ggccagatct
     2101 ggggtctaaa tggctatatt ctgtgccatg gggatttagg aggctcctta actttgctca
     2161 gactcaatac ggtgatcctc ccatatatgt gatggaaaat ggagcatctc aaaaattcca
     2221 ctgtactcaa ttatgtgatg agtggagaat tcaatacctt aaaggataca taaatgaaat
     2281 gctaaaagct ataaaagatg gtgctaatat aaaggggtat acttcctggt ctctgttgga
     2341 taagtttgaa tgggagaaag gatactcaga tagatatgga ttctactatg ttgaatttaa
     2401 cgacagaaat aagcctcgct atccaaaggc ttcagttcaa tattacaaga agattatcat
     2461 tgccaatggg tttcccaatc caagagaggt ggaaagttgg tacctcaaag ctttggaaac
     2521 ttgctctatc aacaatcaga tgcttgctgc agagcccttg ctaagtcaca tgcaaatggt
     2581 tacggagatc gtggtaccca ctgtctgctc cctctgtgtc ctcatcactg ctgttctact
     2641 aatgctcctc ctgaggaggc agagcggagg agggggaagc ccgcgggccc gggatccacc
     2701 ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     2761 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     2821 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     2881 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     2941 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     3001 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     3061 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     3121 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     3181 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     3241 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     3301 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     3361 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     3421 gtacaagtaa agcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     3481 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3541 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3601 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3661 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     3721 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3781 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3841 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3901 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3961 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4021 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4081 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4141 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4201 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4261 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4321 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4381 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4441 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4501 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4561 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4621 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4681 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4741 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4801 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4861 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4921 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4981 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5041 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5101 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5161 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5221 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5281 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5341 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5401 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5461 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5521 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5581 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5641 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5701 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5761 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5821 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5881 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5941 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6001 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6061 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6121 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6181 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6241 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6301 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6361 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6421 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6481 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6541 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6601 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6661 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6721 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6781 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6841 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6901 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6961 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7021 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7081 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7141 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7201 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7261 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7321 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7381 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7441 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7501 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7561 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7621 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7681 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7741 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7801 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7861 ccgaaaagtg ccacctgacg t
//
