LOCUS       V027377                 8867 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027377
VERSION     V027377
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             1660..4239
                     /label="TGFBRAP1(NM_004257)"
                     /note="TGFBRAP1(NM_004257)"
                     /gene="TGFBRAP1"
     CDS             4246..4272
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4276..4302
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4306..4332
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    4434..4658
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4704..5132
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5146..5475
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5542..6336
                     /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    6510..6631
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6680..6696)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6704..6720
                     /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(6728..6758)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6773..6794
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7082..7667)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7838..8698)
                     /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(8699..8803)
                     /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 agtaagctta tgatgagcat caaagccttt
     1681 acgcttgtct ctgctgtgga gcgggagctg ctgatgggcg acaaggagcg cgtcaacata
     1741 gagtgcgtgg agtgctgcgg cagggacctc tacgtgggca ccaacgactg cttcgtctac
     1801 cacttcctgt tggaggagag gccagtgcct gctgggccag ccacgttcac tgccaccaaa
     1861 cagctgcaga gacacttggg cttcaagaag cccgtgaacg agctgcgtgc ggcctcagca
     1921 ctcaacaggc tgctggtgct gtgtgacaac tccatcagcc tggtcaacat gctgaacctc
     1981 gagccagtgc cttcgggggc ccgcatcaag ggggcagcca cgtttgcact gaacgagaac
     2041 cctgtgagtg gggacccctt ctgtgtagaa gtttgcatca tctctgtcaa acgcagaacc
     2101 atccagatgt ttctggtgta cgaggaccgg gtgcagatcg tcaaggaggt gtcgactgcc
     2161 gagcagcccc tcgctgtggc tgtggacggc cacttcctgt gtctggctct gaccactcag
     2221 tacatcatcc acaattacag cacaggcgtc tcccaggacc tgtttcccta ctgcagtgag
     2281 gagaggccgc cgatcgtcaa gaggataggg agacaggagt tcctgctggc gggccccgga
     2341 gggctgggca tgtttgccac agtcgcaggg atatcccagc gcgcccccgt gcactggtcg
     2401 gagaatgtga ttggggcggc tgtgtccttt ccatacgtca tagcgctcga tgacgaattc
     2461 atcacagtcc acagcatgtt ggatcagcaa cagaagcaga cgctgccctt taaggagggc
     2521 catatcctac aggactttga aggaagagtg atcgttgcca caagtaaagg agtttacatc
     2581 ttggttccat tacctttgga aaaacaaata caggatcttc tagcaagccg cagagtagaa
     2641 gaggctttgg ttttagcaaa aggagcccgg aggaacattc caaaggaaaa atttcaggta
     2701 atgtacagaa ggattctgca gcaggcggga tttatacagt ttgcacaact tcagttcctg
     2761 gaagctaaag agctcttcag aagcggccag cttgatgtcc gggagctgat ctctctctac
     2821 cccttcctgt tgcccacctc ctcctccttc acccggtccc accctcctct tcatgagtac
     2881 gcagacctga accagctgac ccagggggac caggagaaga tggccaagtg caaacgcttc
     2941 ctcatgagct acctgaacga ggtccgcagc acagaggtag caaatggcta caaggaggac
     3001 atcgacacag ccttgctcaa actgtatgca gaggctgacc acgacagcct gctggacctc
     3061 ctggtcactg agaacttctg tcttctgacg gacagtgctg cctggctaga gaagcacaaa
     3121 aagtattttg cacttggact gctctatcat tataataacc aagatgctgc tgcagttcag
     3181 ttgtgggtga acattgtgaa tggcgatgtc caggactcca cacgctcaga cctgtatgaa
     3241 tacatcgtgg attttcttac ctactgctta gacgaggaac tagtgtgggc ctatgctgat
     3301 tgggtcctgc agaaaagtga agaggtcgga gttcaggttt tcaccaagag acctttggat
     3361 gaacagcaga agaacagttt taatccagac gacattatca attgccttaa aaaataccct
     3421 aaagcccttg tgaagtatct ggaacatctt gtgatagaca agagactgca gaaagaagag
     3481 tatcacaccc acttagctgt gctgtacctg gaagaggtgc tgctgcagag ggcctccgcc
     3541 agtggcaagg gtgcagaggc caccgagacg caggccaagc tgcggcggct gctccagaaa
     3601 tctgatttat accgagtcca ctttcttctc gagaggctgc agggagctgg cctgcccatg
     3661 gagagcgcca tcctgcacgg gaagctgggc gagcatgaga aggcgctgca tatcctggtg
     3721 cacgagctgc aggactttgc agcggccgag gactactgcc tgtggtgctc cgagggccga
     3781 gacccacccc accgccagca actctttcac acgctgctgg ccatctacct gcatgctggc
     3841 cccactgccc acgagctggc cgtggctgcc gtggacctgc tgaaccgcca cgccaccgaa
     3901 tttgatgcag cccaggtgct gcagatgctg cctgacacct ggtcagtgca gctcctctgc
     3961 ccattcctga tgggggccat gagggacagc atccatgcca ggaggaccat gcaggtggct
     4021 ctcggcctgg ccaggtccga aaacttaatc tacacctacg ataagatgaa gttgaaagga
     4081 agctcaatcc aactctcaga caaaaagctt tgtcagatat gccaaaatcc cttttgtgag
     4141 cctgtgtttg ttagataccc aaatggtggt cttgtgcaca cccactgtgc cgccagcaga
     4201 cacacaaacc ccagctcatc cagtcctggc actcggactg gtacctaccc atacgacgta
     4261 cctgattatg caacttatcc ttatgatgta cctgattatg ctagctatcc ttatgatgtc
     4321 ccagactacg cataggatcc actagtccag tgtggtggaa ttctgcagat atccagcaca
     4381 gtggcggccg ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     4441 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     4501 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     4561 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     4621 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     4681 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4741 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4801 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4861 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4921 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4981 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     5041 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     5101 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     5161 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5221 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5281 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     5341 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5401 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5461 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     5521 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     5581 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     5641 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     5701 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     5761 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5821 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5881 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5941 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     6001 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     6061 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     6121 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     6181 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     6241 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     6301 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     6361 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     6421 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     6481 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     6541 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     6601 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     6661 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6721 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6781 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6841 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6901 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6961 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     7021 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     7081 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     7141 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     7201 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7261 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7321 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7381 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7441 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7501 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     7561 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt
     7621 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     7681 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     7741 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     7801 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     7861 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     7921 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     7981 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     8041 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     8101 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     8161 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     8221 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     8281 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     8341 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     8401 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     8461 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     8521 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     8581 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     8641 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     8701 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     8761 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     8821 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggt
//
