LOCUS       V022171                 8761 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022171
VERSION     V022171
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             1666..4248
                     /label="PIWIL1(NM_004764)"
                     /note="PIWIL1(NM_004764)"
                     /gene="PIWIL1"
     polyA_signal    4328..4552
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4598..5026
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5040..5369
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5436..6230
                     /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    6404..6525
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6574..6590)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6598..6614
                     /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(6622..6652)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6667..6688
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6976..7561)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7732..8592)
                     /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(8593..8697)
                     /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 tccggactca gatctcgagg cggcggaggg
     1621 agtggaggag ggggaagcgg aggcggaggg agtaagcttg gtaccatgac tgggagagcc
     1681 cgagccagag ccagaggaag ggcccgcggt caggagacag cgcagctggt gggctccact
     1741 gccagtcagc aacctggtta tattcagcct aggcctcagc cgccaccagc agagggggaa
     1801 ttatttggcc gtggacggca gagaggaaca gcaggaggaa cagccaagtc acaaggactc
     1861 cagatatctg ctggatttca ggagttatcg ttagcagaga gaggaggtcg tcgtagagat
     1921 tttcatgatc ttggtgtgaa tacaaggcag aacctagacc atgttaaaga atcaaaaaca
     1981 ggttcttcag gcattatagt aaggttaagc actaaccatt tccggctgac atcccgtccc
     2041 cagtgggcct tatatcagta tcacattgac tataacccac tgatggaagc cagaagactc
     2101 cgttcagctc ttctttttca acacgaagat ctaattggaa agtgtcatgc ttttgatgga
     2161 acgatattat ttttacctaa aagactacag caaaaggtta ctgaagtttt tagtaagacc
     2221 cggaatggag aggatgtgag gataacgatc actttaacaa atgaacttcc acctacatca
     2281 ccaacttgtt tgcagttcta taatattatt ttcaggaggc ttttgaaaat catgaatttg
     2341 caacaaattg gacgaaatta ttataaccca aatgacccaa ttgatattcc aagtcacagg
     2401 ttggtgattt ggcctggctt cactacttcc atccttcagt atgaaaacag catcatgctc
     2461 tgcactgacg ttagccataa agtccttcga agtgagactg ttttggattt catgttcaac
     2521 ttttatcatc agacagaaga acataaattt caagaacaag tttccaaaga actaataggt
     2581 ttagttgttc ttaccaagta taacaataag acatacagag tggatgatat tgactgggac
     2641 cagaatccca agagcacctt taagaaagcc gacggctctg aagtcagctt cttagaatac
     2701 tacaggaagc aatacaacca agagatcacc gacttgaagc agcctgtctt ggtcagccag
     2761 cccaagagaa ggcggggccc tggggggaca ctgccagggc ctgccatgct cattcctgag
     2821 ctctgctatc ttacaggtct aactgataaa atgcgtaatg attttaacgt gatgaaagac
     2881 ttagccgttc atacaagact aactccagag caaaggcagc gtgaagtggg acgactcatt
     2941 gattacattc ataaaaacga taatgttcaa agggagcttc gagactgggg tttgagcttt
     3001 gattccaact tactgtcctt ctcaggaaga attttgcaaa cagaaaagat tcaccaaggt
     3061 ggaaaaacat ttgattacaa tccacaattt gcagattggt ccaaagaaac aagaggtgca
     3121 ccattaatta gtgttaagcc actagataac tggctgttga tctatacgcg aagaaattat
     3181 gaagcagcca attcattgat acaaaatcta tttaaagtta caccagccat gggcatgcaa
     3241 atgagaaaag caataatgat tgaagtggat gacagaactg aagcctactt aagagtctta
     3301 cagcaaaagg tcacagcaga cacccagata gttgtctgtc tgttgtcaag taatcggaag
     3361 gacaaatacg atgctattaa aaaatacctg tgtacagatt gccctacccc aagtcagtgt
     3421 gtggtggccc gaaccttagg caaacagcaa actgtcatgg ccattgctac aaagattgcc
     3481 ctacagatga actgcaagat gggaggagag ctctggaggg tggacatccc cctgaagctc
     3541 gtgatgatcg ttggcatcga ttgttaccat gacatgacag ctgggcggag gtcaatcgca
     3601 ggatttgttg ccagcatcaa tgaagggatg acccgctggt tctcacgctg catatttcag
     3661 gatagaggac aggagctggt agatgggctc aaagtctgcc tgcaagcggc tctgagggct
     3721 tggaatagct gcaatgagta catgcccagc cggatcatcg tgtaccgcga tggcgtagga
     3781 gacggccagc tgaaaacact ggtgaactac gaagtgccac agtttttgga ttgtctaaaa
     3841 tccattggta gaggttacaa ccctagacta acggtaattg tggtgaagaa aagagtgaac
     3901 accagatttt ttgctcagtc tggaggaaga cttcagaatc cacttcctgg aacagttatt
     3961 gatgtagagg ttaccagacc agaatggtat gactttttta tcgtgagcca ggctgtgaga
     4021 agtggtagtg tttctcccac acattacaat gtcatctatg acaacagcgg cctgaagcca
     4081 gaccacatac agcgcttgac ctacaagctg tgccacatct attacaactg gccaggtgtc
     4141 attcgtgttc ctgctccttg ccagtacgcc cacaagctgg cttttcttgt tggccagagt
     4201 attcacagag agccaaatct gtcactgtca aaccgccttt actacctcta agaattctgc
     4261 agatatccag cacagtggcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca
     4321 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     4381 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     4441 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     4501 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     4561 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     4621 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4681 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4741 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4801 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4861 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4921 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4981 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     5041 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     5101 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     5161 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     5221 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     5281 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     5341 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     5401 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     5461 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     5521 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     5581 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     5641 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     5701 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     5761 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5821 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5881 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5941 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     6001 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     6061 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     6121 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     6181 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     6241 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     6301 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     6361 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     6421 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     6481 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     6541 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     6601 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     6661 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6721 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6781 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6841 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6901 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6961 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     7021 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     7081 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     7141 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     7201 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     7261 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     7321 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     7381 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     7441 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     7501 cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7561 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7621 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7681 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7741 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7801 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7861 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7921 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7981 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     8041 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     8101 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     8161 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     8221 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     8281 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     8341 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     8401 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8461 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8521 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8581 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8641 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8701 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     8761 t
//
