LOCUS       V030116                 8395 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030116
VERSION     V030116
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             874..1590
                     /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             1642..3885
                     /label="TAP1(NM_000593)"
                     /note="TAP1(NM_000593)"
                     /gene="TAP1"
     polyA_signal    3962..4186
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4232..4660
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4674..5003
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5070..5864
                     /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    6038..6159
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6208..6224)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6232..6248
                     /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(6256..6286)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6301..6322
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6610..7195)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7366..8226)
                     /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(8227..8331)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgcc accatggtga gcaagggcga ggagctgttc
      901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1561 atcactctcg gcatggacga gctgtacaag ggaggcggag gtagtggagg agggggaagc
     1621 ggaggcggag ggagtgaatt catggctagc tctaggtgtc ccgctccccg cgggtgccgc
     1681 tgcctccccg gagcttctct cgcatggctg gggacagtac tgctacttct cgccgactgg
     1741 gtgctgctcc ggaccgcgct gccccgcata ttctccctgc tggtgcccac cgcgctgcca
     1801 ctgctccggg tctgggcggt gggcctgagc cgctgggccg tgctctggct gggggcctgc
     1861 ggggtcctca gggcaacggt tggctccaag agcgaaaacg caggtgccca gggctggctg
     1921 gctgctttga agccattagc tgcggcactg ggcttggccc tgccgggact tgccttgttc
     1981 cgagagctga tctcatgggg agcccccggg tccgcggata gcaccaggct actgcactgg
     2041 ggaagtcacc ctaccgcctt cgttgtcagt tatgcagcgg cactgcccgc agcagccctg
     2101 tggcacaaac tcgggagcct ctgggtgccc ggcggtcagg gcggctctgg aaaccctgtg
     2161 cgtcggcttc taggctgcct gggctcggag acgcgccgcc tctcgctgtt cctggtcctg
     2221 gtggtcctct cctctcttgg ggagatggcc attccattct ttacgggccg cctcactgac
     2281 tggattctac aagatggctc agccgatacc ttcactcgaa acttaactct catgtccatt
     2341 ctcaccatag ccagtgcagt gctggagttc gtgggtgacg ggatctataa caacaccatg
     2401 ggccacgtgc acagccactt gcagggagag gtgtttgggg ctgtcctgcg ccaggagacg
     2461 gagtttttcc aacagaacca gacaggtaac atcatgtctc gggtaacaga ggacacgtcc
     2521 accctgagtg attctctgag tgagaatctg agcttatttc tgtggtacct ggtgcgaggc
     2581 ctatgtctct tggggatcat gctctgggga tcagtgtccc tcaccatggt caccctgatc
     2641 accctgcctc tgcttttcct tctgcccaag aaggtgggaa aatggtacca gttgctggaa
     2701 gtgcaggtgc gggaatctct ggcaaagtcc agccaggtgg ccattgaggc tctgtcggcc
     2761 atgcctacag ttcgaagctt tgccaacgag gagggcgaag cccagaagtt tagggaaaag
     2821 ctgcaagaaa taaagacact caaccagaag gaggctgtgg cctatgcagt caactcctgg
     2881 accactagta tttcaggtat gctgctgaaa gtgggaatcc tctacattgg tgggcagctg
     2941 gtgaccagtg gggctgtaag cagtgggaac cttgtcacat ttgttctcta ccagatgcag
     3001 ttcacccagg ctgtggaggt actgctctcc atctacccca gagtacagaa ggctgtgggc
     3061 tcctcagaga aaatatttga gtacctggac cgcacccctc gctgcccacc cagtggtctg
     3121 ttgactccct tacacttgga gggccttgtc cagttccaag atgtctcctt tgcctaccca
     3181 aaccgcccag atgtcttagt gctacagggg ctgacattca ccctacgccc tggcgaggtg
     3241 acggcgctgg tgggacccaa tgggtctggg aagagcacag tggctgccct gctgcagaat
     3301 ctgtaccagc ccaccggggg acagctgctg ttggatggga agccccttcc ccaatatgag
     3361 caccgctacc tgcacaggca ggtggctgca gtgggacaag agccacaggt atttggaaga
     3421 agtcttcaag aaaatattgc ctatggcctg acccagaagc caactatgga ggaaatcaca
     3481 gctgctgcag taaagtctgg ggcccatagt ttcatctctg gactccctca gggctatgac
     3541 acagaggtag acgaggctgg gagccagctg tcagggggtc agcgacaggc agtggcgttg
     3601 gcccgagcat tgatccggaa accgtgtgta cttatcctgg atgatgccac cagtgccctg
     3661 gatgcaaaca gccagttaca ggtggagcag ctcctgtacg aaagccctga gcggtactcc
     3721 cgctcagtgc ttctcatcac ccagcacctc agcctggtgg agcaggctga ccacatcctc
     3781 tttctggaag gaggcgctat ccgggagggg ggaacccacc agcagctcat ggagaaaaag
     3841 gggtgctact gggccatggt gcaggctcct gcagatgctc cagaatgagg atcctgatat
     3901 ccagcacagt ggcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     3961 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     4021 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     4081 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     4141 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     4201 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     4261 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4321 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4381 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4441 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4501 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4561 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4621 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4681 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4741 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4801 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4861 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4921 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4981 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     5041 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     5101 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     5161 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     5221 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     5281 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     5341 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     5401 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5461 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5521 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5581 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5641 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5701 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5761 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5821 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5881 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5941 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     6001 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     6061 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     6121 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     6181 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6241 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6301 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6361 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6421 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6481 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6541 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6601 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6661 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6721 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6781 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6841 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6901 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6961 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7021 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     7081 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7141 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7201 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7261 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7321 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7381 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7441 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7501 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7561 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7621 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7681 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7741 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7801 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7861 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7921 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7981 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8041 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8101 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8161 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8221 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8281 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8341 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggt
//
