LOCUS       V032536                 8902 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032536
VERSION     V032536
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..4392
                     /label="HK1(NM_000188)"
                     /note="HK1(NM_000188)"
                     /gene="HK1"
     polyA_signal    4469..4693
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4739..5167
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5181..5510
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5577..6371
                     /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    6545..6666
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6715..6731)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6739..6755
                     /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(6763..6793)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6808..6829
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7117..7702)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7873..8733)
                     /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(8734..8838)
                     /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 catgatcgcc gcgcagctcc tggcctatta cttcacggag
     1681 ctgaaggatg accaggtcaa aaagattgac aagtatctct atgccatgcg gctctccgat
     1741 gaaactctca tagatatcat gactcgcttc aggaaggaga tgaagaatgg cctctcccgg
     1801 gattttaatc caacagccac agtcaagatg ttgccaacat tcgtaaggtc cattcctgat
     1861 ggctctgaaa agggagattt cattgccctg gatcttggtg ggtcttcctt tcgaattctg
     1921 cgggtgcaag tgaatcatga gaaaaaccag aatgttcaca tggagtccga ggtttatgac
     1981 accccagaga acatcgtgca cggcagtgga agccagcttt ttgatcatgt tgctgagtgc
     2041 ctgggagatt tcatggagaa aaggaagatc aaggacaaga agttacctgt gggattcacg
     2101 ttttcttttc cttgccaaca atccaaaata gatgaggcca tcctgatcac ctggacaaag
     2161 cgatttaaag cgagcggagt ggaaggagca gatgtggtca aactgcttaa caaagccatc
     2221 aaaaagcgag gggactatga tgccaacatc gtagctgtgg tgaatgacac agtgggcacc
     2281 atgatgacct gtggctatga cgaccagcac tgtgaagtcg gcctgatcat cggcactggc
     2341 accaatgctt gctacatgga ggaactgagg cacattgatc tggtggaagg agacgagggg
     2401 aggatgtgta tcaatacaga atggggagcc tttggagacg atggatcatt agaagacatc
     2461 cggacagagt ttgacaggga gatagaccgg ggatccctca accctggaaa acagctgttt
     2521 gagaagatgg tcagtggcat gtacttggga gagctggttc gactgatcct agtcaagatg
     2581 gccaaggagg gcctcttatt tgaagggcgg atcaccccgg agctgctcac ccgagggaag
     2641 tttaacacca gtgatgtgtc agccatcgaa aagaataagg aaggcctcca caatgccaaa
     2701 gaaatcctga cccgcctggg agtggagccg tccgatgatg actgtgtctc agtccagcac
     2761 gtttgcacca ttgtctcatt tcgctcagcc aacttggtgg ctgccacact gggcgccatc
     2821 ttgaaccgcc tgcgtgataa caagggcaca cccaggctgc ggaccacggt tggtgtcgac
     2881 ggatctcttt acaagacgca cccacagtat tcccggcgtt tccacaagac tctaaggcgc
     2941 ttggtgccag actccgatgt gcgcttcctc ctctcggaga gtggcagcgg caagggggct
     3001 gccatggtga cggcggtggc ctaccgcttg gccgagcagc accggcagat agaggagacc
     3061 ctggctcatt tccacctcac caaggacatg ctgctggagg tgaagaagag gatgcgggcc
     3121 gagatggagc tggggctgag gaagcagacg cacaacaatg ccgtggttaa gatgctgccc
     3181 tccttcgtcc ggagaactcc cgacgggacc gagaatggtg acttcttggc cctggatctt
     3241 ggaggaacca atttccgtgt gctgctggtg aaaatccgta gtgggaaaaa gagaacggtg
     3301 gaaatgcaca acaagatcta cgccattcct attgaaatca tgcagggcac tggggaagag
     3361 ctgtttgatc acattgtctc ctgcatctct gacttcttgg actacatggg gatcaaaggc
     3421 cccaggatgc ctctgggctt cacgttctca tttccctgcc agcagacgag tctggacgcg
     3481 ggaatcttga tcacgtggac aaagggtttt aaggcaacag actgcgtggg ccacgatgta
     3541 gtcaccttac taagggatgc gataaaaagg agagaggaat ttgacctgga cgtggtggct
     3601 gtggtcaacg acacagtggg caccatgatg acctgtgctt atgaggagcc cacctgtgag
     3661 gttggactca ttgttgggac cggcagcaat gcctgctaca tggaggagat gaagaacgtg
     3721 gagatggtgg agggggacca ggggcagatg tgcatcaaca tggagtgggg ggcctttggg
     3781 gacaacgggt gtctggatga tatcaggaca cactacgaca gactggtgga cgaatattcc
     3841 ctaaatgctg ggaaacaaag gtatgagaag atgatcagtg gtatgtacct gggtgaaatc
     3901 gtccgcaaca tcttaatcga cttcaccaag aagggattcc tcttccgagg gcagatctct
     3961 gagacgctga agacccgggg catctttgag accaagtttc tctctcagat cgagagtgac
     4021 cgattagcac tgctccaggt ccgggctatc ctccagcagc taggtctgaa tagcacctgc
     4081 gatgacagta tcctcgtcaa gacagtgtgc ggggtggtgt ccaggagggc cgcacagctg
     4141 tgtggcgcag gcatggctgc ggttgtggat aagatccgcg agaacagagg actggaccgt
     4201 ctgaatgtga ctgtgggagt ggacgggaca ctctacaagc ttcatccaca cttctccaga
     4261 atcatgcacc agacggtgaa ggaactgtca ccaaaatgta acgtgtcctt cctcctgtct
     4321 gaggatggca gcggcaaggg ggccgccctc atcacggccg tgggcgtgcg gttacgcaca
     4381 gaggcaagca gctaaggatc ctgatatcca gcacagtggc ggccgctcga gtctagaggg
     4441 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     4501 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     4561 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     4621 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     4681 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     4741 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4801 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4861 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4921 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4981 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5041 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5101 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5161 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     5221 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     5281 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     5341 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5401 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     5461 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     5521 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     5581 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     5641 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     5701 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     5761 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     5821 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     5881 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     5941 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     6001 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     6061 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     6121 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     6181 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     6241 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     6301 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     6361 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     6421 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     6481 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     6541 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6601 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6661 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     6721 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6781 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6841 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6901 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6961 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     7021 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     7081 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     7141 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7201 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7261 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7321 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7381 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7441 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7501 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     7561 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7621 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac
     7681 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7741 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7801 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7861 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7921 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7981 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     8041 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     8101 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     8161 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     8221 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8281 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8341 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8401 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8461 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8521 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8581 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8641 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8701 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8761 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8821 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     8881 agatctcccg atcccctatg gt
//
