LOCUS       V044990                 7639 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044990
VERSION     V044990
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..3129
                     /label="Trim30a(NM_009099)"
                     /note="Trim30a(NM_009099)"
                     /gene="Trim30a"
     polyA_signal    3206..3430
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3476..3904
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3918..4247
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4314..5108
                     /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    5282..5403
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5452..5468)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5476..5492
                     /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(5500..5530)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5545..5566
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5854..6439)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6610..7470)
                     /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(7471..7575)
                     /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 catggcctca tcagtcctgg agatgataaa ggaggaagta
     1681 acctgtccta tctgtttgga gctcctgaag gaacctgtga gtgctgattg taaccacagc
     1741 ttctgcagag cctgcatcac actgaattat gagtccaaca gaaacacaga cgggaagggc
     1801 aactgccctg tatgccgagt tccttaccca tttgggaatc tgaggcctaa tctacatgtg
     1861 gccaacatag tagagaggct caagggattc aagtccattc cagaggagga gcagaaggtg
     1921 aatatctgtg cacaacatgg agagaaactc cggctcttct gtaggaagga catgatggtc
     1981 atctgctggc tttgtgagcg atctcaggag caccgtggtc accaaacagc tctcattgaa
     2041 gaggttgacc aagaatacaa ggagaagctg cagggagctc tgtggaagct gatgaaaaag
     2101 gcaaaaatat gtgatgaatg gcaggatgac cttcaactgc agagagttga ctgggagaac
     2161 caaatacaga tcaatgtaga aaatgttcag agacagttta aaggactaag agacctcctg
     2221 gactccaagg agaatgagga gctgcagaag ctgaagaaag agaagaaaga ggttatggaa
     2281 aagctggaag agtctgaaaa tgagctggag gatcagacag agttggtgag agacctcatc
     2341 tcagatgtgg aacatcattt ggagctctca accttagaaa tgctgcaggg tgcaaattgt
     2401 gtcctgagaa ggagtcagtc cttaagcctg caacagcccc aaactgtccc ccaaaagaga
     2461 aaaagaacat tccaagctcc agatctgaaa ggcatgctgc aagtgtatca aggactcatg
     2521 gatatccagc aatactgggt tcatatgact ctacatgcaa ggaacaatgc agtcattgcc
     2581 attaacaaag aaaaaagaca aatacagtat agaagttaca atacggttcc agtttctgag
     2641 atctaccatt tgggtgtcct gggatatcca gctctttcct cagggaagca ttactgggaa
     2701 gtagacatat ctagaagtga tgcctggctc ctcggattaa atgacggaaa gtgtgctcaa
     2761 ccccaacttc actcaaagga agaaatgggc atcaaaaaaa accttcattc tcagatcaaa
     2821 caaaatgtat tgtttcagcc taaatgtggc tactgggtta tagggatgaa gaatccgtct
     2881 gtatacaagg cctttgatga gtgttctatc acccacaatt ccagtatcct ggtcatctct
     2941 ctgcctgatc gtcccagtcg tgtcggagtt ttcctggatc ggaaagctgg cactctctca
     3001 ttttatgatg tttctaactg cggtgctctc atctataggt tctatgaccc tgccttccct
     3061 gttgaagtct atccatattt taatcctatg aaatgttcag agccaatgac tatatgcggg
     3121 ccaccctcct aaggatcctg atatccagca cagtggcggc cgctcgagtc tagagggccc
     3181 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3241 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3301 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3361 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     3421 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     3481 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3541 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3601 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3661 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3721 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3781 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3841 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3901 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3961 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4021 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4081 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4141 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4201 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     4261 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     4321 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4381 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4441 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4501 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4561 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4621 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4681 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4741 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4801 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     4861 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4921 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4981 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5041 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5101 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     5161 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5221 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5281 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5341 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5401 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     5461 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5521 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5581 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5641 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5701 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5761 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5821 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5881 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5941 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6001 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6061 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6121 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6181 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6241 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6301 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6361 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     6421 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6481 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6541 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6601 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6661 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6721 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6781 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6841 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6901 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6961 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7021 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7081 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7141 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7201 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7261 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7321 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7381 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7441 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7501 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7561 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     7621 tctcccgatc ccctatggt
//
