LOCUS       V028684                 8464 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028684
VERSION     V028684
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..3954
                     /label="TRIM2(NM_015271)"
                     /note="TRIM2(NM_015271)"
                     /gene="TRIM2"
     polyA_signal    4031..4255
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4301..4729
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4743..5072
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5139..5933
                     /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    6107..6228
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6277..6293)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6301..6317
                     /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(6325..6355)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6370..6391
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6679..7264)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7435..8295)
                     /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(8296..8400)
                     /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 catgcacagg agtggccgtt atggaacgca gcagcagcgt
     1681 gcagggtcaa agacagccgg ccccccatgt cagtggtcta ggatggccag tgaaggcacc
     1741 aacatcccaa gtcctgtggt gcgccagatt gacaagcagt ttctgatttg cagtatatgc
     1801 ctggaacggt acaagaatcc caaggttctc ccctgtctgc acactttctg cgagaggtgc
     1861 ctgcagaact acattcctgc ccacagttta accctctcct gcccagtgtg ccgccagacc
     1921 tccatcctgc ccgagaaagg ggtggccgcg ctccagaaca atttcttcat cacaaacctg
     1981 atggacgtgc tgcagcgaac tccaggcagc aacgctgagg agtcttccat cctggagaca
     2041 gtcactgctg tggctgcggg aaagcctctc tcttgcccaa accacgatgg gaatgtgatg
     2101 gaattttact gccagtcctg tgagactgcc atgtgtcggg agtgcacgga gggggagcac
     2161 gcagagcacc ccacagttcc actcaaggat gtggtggaac agcacaaggc ctcgctccag
     2221 gtccagctgg atgctgtcaa caaaaggctc ccagaaatag attctgctct tcagttcatc
     2281 tctgaaatca ttcatcagtt aaccaaccaa aaggccagca tcgtggatga cattcattcc
     2341 acctttgatg agctccagaa gactttaaat gtgcgcaaga gtgtgctgct tatggaattg
     2401 gaggtcaact atggcctcaa acacaaagtc ctccagtcgc agctggatac tctgctccag
     2461 gggcaggaga gcattaagag ctgcagcaac ttcacagcgc aggccctcaa ccatggcacg
     2521 gagaccgagg tcctactggt gaagaagcag atgagcgaga agctgaacga gctggccgac
     2581 caggacttcc ccttgcaccc gcgggagaac gaccagctgg atttcatcgt ggaaaccgag
     2641 gggctgaaga agtccatcca caacctcggg acgatcttaa ccaccaacgc cgttgcctca
     2701 gagacagtgg ccacgggcga ggggctgcgg cagaccatca tcgggcagcc catgtccgtc
     2761 accatcacca ccaaggacaa agacggtgag ctgtgcaaaa ccggcaacgc ctacctcacc
     2821 gccgaactga gcacccccga cgggagcgtg gcagacgggg agatcctgga caacaagaac
     2881 ggcacctatg agtttttgta cactgtccag aaggaagggg actttaccct gtctctgaga
     2941 ctctatgacc agcacatccg aggcagcccg tttaagctga aagtgatccg atccgctgat
     3001 gtgtctccca ccacagaagg cgtgaagagg cgcgttaagt ccccggggag cggccacgtc
     3061 aagcagaaag ctgtgaaaag acccgcaagc atgtacagca ctggaaaacg aaaagagaat
     3121 cccatcgaag acgatttgat ctttcgagtg ggtaccaaag gaagaaataa aggagagttt
     3181 acaaatcttc agggggtagc tgcatctaca aatggaaaga tattaattgc agacagtaac
     3241 aaccaatgtg tgcagatatt ttccaatgat ggccagttca aaagtcgttt tggcatacgg
     3301 ggacgctctc cggggcagct gcagcggccc acaggagtgg ctgtacatcc cagtggggac
     3361 ataatcattg ccgattatga taataaatgg gtcagcattt tctcctccga tgggaaattt
     3421 aagacaaaaa ttggatcagg aaagctgatg ggacccaaag gagtttctgt ggaccgcaat
     3481 gggcacatta ttgttgtgga caacaaggcg tgctgcgtgt ttatcttcca gccaaacggg
     3541 aaaatagtca ccaggtttgg tagccgagga aatggggaca ggcagtttgc aggtccccat
     3601 tttgcagctg taaatagcaa taatgagatt attattacag atttccataa tcattctgtc
     3661 aaggtgttta atcaggaagg agaattcatg ttgaagtttg gctcaaatgg agaaggaaat
     3721 gggcagttta atgctccaac aggtgtagca gtggattcaa atggaaacat cattgtggcc
     3781 gactggggaa acagcaggat ccaggttttt gatgggagtg gatcattttt gtcctacatt
     3841 aacacatctg ctgacccact ctatggcccc caaggcctgg ccctaacttc agatggtcat
     3901 gttgtggttg cagactctgg aaatcactgt ttcaaagtct atcgatactt acagtaagga
     3961 tcctgatatc cagcacagtg gcggccgctc gagtctagag ggcccgttta aacccgctga
     4021 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     4081 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     4141 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     4201 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     4261 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     4321 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4381 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4441 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4501 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4561 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4621 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4681 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     4741 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4801 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4861 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4921 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4981 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     5041 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     5101 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     5161 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     5221 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     5281 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     5341 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     5401 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5461 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5521 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5581 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5641 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     5701 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5761 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5821 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5881 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5941 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     6001 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     6061 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     6121 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6181 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     6241 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     6301 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6361 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6421 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6481 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6541 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6601 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6661 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6721 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6781 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6841 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6901 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6961 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7021 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7081 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     7141 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7201 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     7261 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     7321 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     7381 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7441 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7501 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7561 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7621 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7681 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7741 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7801 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7861 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7921 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7981 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     8041 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     8101 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     8161 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     8221 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     8281 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     8341 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     8401 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     8461 tggt
//
