LOCUS       V033037                 8229 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033037
VERSION     V033037
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..3720
                     /label="TSPYL2(NM_022117)"
                     /note="TSPYL2(NM_022117)"
                     /gene="TSPYL2"
     polyA_signal    3796..4020
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4066..4494
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4508..4837
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4904..5698
                     /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    5872..5993
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6042..6058)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6066..6082
                     /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(6090..6120)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6135..6156
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6444..7029)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7200..8060)
                     /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(8061..8165)
                     /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 catggaccgc ccagatgagg ggcctccggc caagacccgc
     1681 cgcctgagca gctccgagtc tccacagcgc gacccgcccc cgccgccgcc gccgccgccg
     1741 ctcctccgac tgccgctgcc tccaccccag cagcgcccga ggctccagga ggaaacggag
     1801 gcggcacagg tgctggccga tatgaggggg gtgggactgg gccccgcgct gcccccgccg
     1861 cctccctatg tcattctcga ggaggggggg atccgcgcat acttcacgct cggtgctgag
     1921 tgtcccggct gggattctac catcgagtcg gggtatgggg aggcgccccc gcccacggag
     1981 agcctggaag cactccccac tcctgaggcc tcggggggga gcctggaaat cgattttcag
     2041 gttgtacagt cgagcagttt tggtggagag ggggccctag aaacctgtag cgcagtgggg
     2101 tgggcgcccc agaggttagt tgacccgaag agcaaggaag aggcgatcat catagtggag
     2161 gatgaggatg aggatgagcg ggagagtatg aggagcagca ggaggcggcg gcggcggcgg
     2221 aggaggaagc agaggaaggt gaagagggaa agcagagaga gaaatgccga gaggatggag
     2281 agcatcctgc aggcactgga ggatattcag ctggatctgg aggcagtgaa catcaaggca
     2341 ggcaaagcct tcctgcgtct caagcgcaag ttcatccaga tgcgaagacc cttcctggag
     2401 cgcagagacc tcatcatcca gcatatccca ggcttctggg tcaaagcatt cctcaaccac
     2461 cccagaattt caattttgat caaccgacgt gatgaagaca ttttccgcta cttgaccaat
     2521 ctgcaggtac aggatctcag acatatctcc atgggctaca aaatgaagct gtacttccag
     2581 actaacccct acttcacaaa catggtgatt gtcaaggagt tccagcgcaa ccgctcaggc
     2641 cggctggtgt ctcactcaac cccaatccgc tggcaccggg gccaggaacc ccaggcccgt
     2701 cgtcacggga accaggatgc gagccacagc tttttcagct ggttctcaaa ccatagcctc
     2761 ccagaggctg acaggattgc tgagattatc aagaatgatc tgtgggttaa ccctctacgc
     2821 tactacctga gagaaagggg ctccaggata aagagaaaga agcaagaaat gaagaaacgt
     2881 aaaaccaggg gcagatgtga ggtggtgatc atggaagacg cccctgacta ttatgcagtg
     2941 gaagacattt tcagcgagat ctcagacatt gatgagacaa ttcatgacat caagatctct
     3001 gacttcatgg agaccaccga ctacttcgag accactgaca atgagataac tgacatcaat
     3061 gagaacatct gcgacagcga gaatcctgac cacaatgagg tccccaacaa cgagaccact
     3121 gataacaacg agagtgctga tgaccacgaa accactgaca acaatgagag tgcagatgac
     3181 aacaacgaga atcctgaaga caataacaag aacactgatg acaacgaaga gaaccctaac
     3241 aacaacgaga acacttacgg caacaacttc ttcaaaggtg gcttctgggg cagccatggc
     3301 aacaaccagg acagcagcga cagtgacaat gaagcagatg aggccagtga tgatgaagat
     3361 aatgatggca acgaaggtga caatgagggc agtgatgatg atggcaatga aggtgacaat
     3421 gaaggcagcg atgatgacga cagagacatt gagtactatg agaaagttat tgaagacttt
     3481 gacaaggatc aggctgacta cgaggacgtg atagagatca tctcagacga atcagtggaa
     3541 gaagagggca ttgaggaagg catccagcaa gatgaggaca tctatgagga aggaaactat
     3601 gaggaggaag gaagtgaaga tgtctgggaa gaaggggaag attcggacga ctctgaccta
     3661 gaggatgtgc ttcaggtccc aaacggttgg gccaatccgg ggaagagggg gaaaaccgga
     3721 taggatcctg atatccagca cagtggcggc cgctcgagtc tagagggccc gtttaaaccc
     3781 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     3841 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     3901 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     3961 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     4021 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     4081 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4141 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4201 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4261 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     4321 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4381 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     4441 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     4501 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4561 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4621 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4681 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4741 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     4801 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     4861 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     4921 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4981 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     5041 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     5101 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     5161 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     5221 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     5281 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     5341 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     5401 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     5461 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     5521 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     5581 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     5641 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     5701 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     5761 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5821 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     5881 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5941 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     6001 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     6061 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6121 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6181 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6241 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6301 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6361 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6421 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6481 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6541 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6601 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6661 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6721 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6781 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6841 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     6901 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6961 caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     7021 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7081 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7141 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7201 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7261 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7321 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7381 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7441 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7501 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7561 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7621 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7681 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7741 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7801 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7861 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7921 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7981 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     8041 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     8101 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     8161 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     8221 ccctatggt
//
