LOCUS       V020629                 7780 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020629
VERSION     V020629
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             1387..2523
                     /label="TGOLN2(NM_001206844)"
                     /note="TGOLN2(NM_001206844)"
                     /gene="TGOLN2"
     misc_feature    2556..3144
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3210..3258
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3460..3888
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3902..4231
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4298..5092
                     /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    5268..5389
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5438..5454)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5462..5478
                     /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(5486..5516)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5531..5552
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5840..6428)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6599..7459)
                     /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(7460..7564)
                     /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 ggtaccatgc ggttcgtggt tgccttggtc ctcctgaacg tcgcagcggc gggagccgtg
     1441 ccgctcttgg ccaccgaaag cgtcaagcaa gaagaagctg gagtacggcc ttctgcagga
     1501 aacgtctcca cccaccccag cttgagccaa cggcctggag gctctaccaa gtcgcatccg
     1561 gagccgcaga ctccaaaaga cagccctagc aagtcgagtg cggaggcgca gaccccagaa
     1621 gacaccccca acaagtcggg tgcggaggca aagacccaaa aagacagctc caacaagtcg
     1681 ggtgcggagg caaagaccca aaaaggcagc actagcaagt cgggttcgga ggcgcagacc
     1741 acaaaagaca gcactagtaa gtcgcatccg gagctgcaga ctccaaaaga cagcactggc
     1801 aaatcgggtg cggaggcgca gaccccagaa gacagcccca acaggtcggg tgcggaggca
     1861 aagacccaaa aagacagccc tagcaagtca ggttcggagg cgcagaccac aaaagatgtc
     1921 cctaataagt cgggtgcgga cggccagacc ccaaaagacg gctccagcaa gtcgggtgcg
     1981 gaggatcaga ccccaaaaga cgtccctaac aagtcgggtg cggagaagca gactccaaaa
     2041 gacggctcta acaagtccgg tgcagaggag cagggcccaa tagacgggcc cagcaagtcg
     2101 ggtgcggagg agcagacctc aaaagacagc cctaacaagg aggaagttaa gtcttcagag
     2161 cctactgagg atgtggagcc caaagaggct gaagatgatg atacaggacc cgaggagggc
     2221 tcaccgccca aagaagagaa agaaaagatg tccggttctg cctccagtga gaaccgtgaa
     2281 gggacacttt cggattccac gggtagcgag aaggatgacc tttatccgaa cggttctgga
     2341 aatggcagcg cggagagcag ccacttcttt gcatatctgg tgactgcagc cattcttgtg
     2401 gctgtcctct atatcgctca tcacaacaag cggaagatca ttgcttttgt cctggaagga
     2461 aaaagatcta aagtcacccg gcggccaaag gccagtgact accaacgttt ggaccagaag
     2521 tcctaactcg agtctagagg gcccgatatc tcgacaatca acctctggat tacaaaattt
     2581 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     2641 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     2701 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     2761 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     2821 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     2881 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     2941 tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc
     3001 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3061 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3121 tctccctttg ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa cacggaagga
     3181 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     3241 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     3301 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     3361 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     3421 tagcagatct gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     3481 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3541 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3601 aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3661 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3721 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3781 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg
     3841 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     3901 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3961 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4021 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4081 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4141 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     4201 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     4261 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4321 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4381 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4441 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4501 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4561 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4621 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4681 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4741 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4801 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     4861 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4921 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4981 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5041 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5101 ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5161 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5221 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     5281 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5341 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     5401 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5461 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5521 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5581 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5641 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5701 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5761 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5821 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5881 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5941 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6001 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
     6061 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6121 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6181 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6241 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     6301 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6361 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6421 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6481 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6541 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6601 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6661 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6721 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6781 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6841 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6901 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6961 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7021 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7081 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7141 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7201 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7261 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7321 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7381 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7441 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7501 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7561 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     7621 cctatggtcg actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     7681 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     7741 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
