LOCUS       V022549                 9254 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022549
VERSION     V022549
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             908..1624
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="enhanced YFP"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EYFP"
     CDS             1631..4768
                     /label="OGT(NM_181672)"
                     /note="OGT(NM_181672)"
                     /gene="OGT"
     polyA_signal    4821..5045
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5091..5519
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5533..5862
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5929..6723
                     /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    6897..7018
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7067..7083)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7091..7107
                     /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(7115..7145)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7160..7181
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7469..8054)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8225..9085)
                     /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(9086..9190)
                     /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 ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
      961 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     1021 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1081 gcccaccctc gtgaccacct tcggctacgg cctgcagtgc ttcgcccgct accccgacca
     1141 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1201 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1261 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1321 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     1381 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     1441 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1501 caaccactac ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     1561 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1621 caaggaattc atggcgtctt ccgtgggcaa cgtggccgac agcacagaac caacgaaacg
     1681 tatgctttcc ttccaagggt tagctgagtt ggcacatcga gaatatcagg caggagattt
     1741 tgaggcagct gagagacact gcatgcagct ctggagacaa gagccagaca atactggtgt
     1801 gcttttatta ctttcatcta tacacttcca gtgtcgaagg ctggacagat ctgctcactt
     1861 tagcactctg gcaattaaac agaaccccct tctggcagaa gcttattcga atttggggaa
     1921 tgtgtacaag gaaagagggc agttgcagga ggcaattgag cattatcgac atgcattgcg
     1981 tctcaaacct gatttcatcg atggttatat taacctggca gccgccttgg tagcagcggg
     2041 tgacatggaa ggggcagtac aagcttacgt ctctgctctt cagtacaatc ctgatttgta
     2101 ctgtgttcgc agtgacctgg ggaacctgct caaagccctg ggtcgcttgg aagaagccaa
     2161 ggcatgttat ttgaaagcaa ttgagacgca accgaacttt gcagtagctt ggagtaatct
     2221 tggctgtgtt ttcaatgcac aaggggaaat ttggcttgca attcatcact ttgaaaaggc
     2281 tgtcaccctt gacccaaact ttctggatgc ttatatcaat ttaggaaatg tcttgaaaga
     2341 ggcacgcatt tttgacagag ctgtggcagc ttatcttcgt gccctaagtt tgagtccaaa
     2401 tcacgcagtg gtgcacggca acctggcttg tgtatactat gagcaaggcc tgatagatct
     2461 ggcaatagac acctacaggc gggctatcga actacaacca catttccctg atgcttactg
     2521 caacctagcc aatgctctca aagagaaggg cagtgttgct gaagcagaag attgttataa
     2581 tacagctctc cgtctgtgtc ccacccatgc agactctctg aataacctag ccaatatcaa
     2641 acgagaacag ggaaacattg aagaggcagt tcgcttgtat cgtaaagcat tagaagtctt
     2701 cccagagttt gctgctgccc attcaaattt agcaagtgta ctgcagcagc agggaaaact
     2761 gcaggaagct ctgatgcatt ataaggaggc tattcgaatc agtcctacct ttgctgatgc
     2821 ctactctaat atgggaaaca ctctaaagga gatgcaggat gttcagggag ccttgcagtg
     2881 ttatacgcgt gccatccaaa ttaatcctgc atttgcagat gcacatagca atctggcttc
     2941 cattcataag gattcaggga atattccaga agccatagct tcttaccgca cggctctgaa
     3001 acttaagcct gattttcctg atgcttattg taacttggct cattgcctgc agattgtctg
     3061 tgattggaca gactatgatg agcgaatgaa gaagttggtc agtattgtgg ctgaccagtt
     3121 agagaagaat aggttgcctt ctgtgcatcc tcatcatagt atgctatatc ctctttctca
     3181 tggcttcagg aaggctattg ctgagaggca cggcaacctg tgcttagata agattaatgt
     3241 tcttcataaa ccaccatatg aacatccaaa agacttgaag ctcagtgatg gtcggctgcg
     3301 tgtaggatat gtgagttccg actttgggaa tcatcctact tctcacctta tgcagtctat
     3361 tccaggcatg cacaatcctg ataaatttga ggtgttctgt tatgccctga gcccagacga
     3421 tggcacaaac ttccgagtga aggtgatggc agaagccaat catttcattg atctttctca
     3481 gattccatgc aatggaaaag cagctgatcg catccatcag gatggaattc atatccttgt
     3541 aaatatgaat ggctatacta agggcgctcg aaatgagctt tttgctctca ggccagctcc
     3601 tattcaggca atgtggctgg gataccctgg gacgagtggt gcgcttttca tggattatat
     3661 tatcactgat caggaaactt cgccagctga agttgctgag cagtattccg agaaattggc
     3721 ttatatgccc cacacttttt ttattggtga tcatgctaat atgttccctc acctgaagaa
     3781 aaaagcagtc atcgatttta agtccaatgg gcacatttat gacaatcgga tagttctgaa
     3841 tggcatcgac ctcaaagcat ttcttgatag tctaccagat gtgaaaattg tcaagatgaa
     3901 gtgtcctgat ggaggagaca atgcagatag cagtaacaca gctcttaata tgcctgttat
     3961 tcctatgaat actattgcag aagcagttat tgaaatgatt aaccgaggac agattcaaat
     4021 aacaattaat ggattcagta ttagcaatgg actggcaact actcagatca acaataaggc
     4081 tgcaactgga gaggaggttc cccgtaccat tattgtaacc acccgttctc agtacgggtt
     4141 accagaagat gccatcgtat actgtaactt taatcagttg tataaaattg acccttctac
     4201 tttgcagatg tgggcaaaca ttctgaagcg tgttcccaat agtgtactct ggctgttgcg
     4261 ttttccagca gtaggagaac ctaatattca acagtatgca caaaacatgg gcctgcccca
     4321 gaaccgtatc attttttcac ctgttgctcc taaagaggaa cacgtcagga gaggccagct
     4381 ggctgatgtc tgcttggaca ctccactctg taatgggcac accacaggga tggatgtcct
     4441 ctgggcaggg acccccatgg tgactatgcc aggagagact cttgcttctc gagttgcagc
     4501 atcccagctc acttgcttag gttgtcttga gcttattgct aaaaacagac aagaatatga
     4561 agacatagct gtgaagctgg gaactgatct agaatacctg aagaaagttc gtggcaaagt
     4621 ctggaagcaa agaatatcta gccctctgtt caacaccaaa caatacacaa tggaactaga
     4681 gcggctctat ctacagatgt gggagcatta tgcagctggc aacaaacctg accacatgat
     4741 taagcctgtt gaagtcactg agtcagcaac cggttagctc gagtctagag ggcccgttta
     4801 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     4861 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     4921 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     4981 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     5041 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     5101 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     5161 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     5221 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     5281 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     5341 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     5401 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     5461 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     5521 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     5581 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     5641 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     5701 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     5761 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     5821 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     5881 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     5941 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     6001 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     6061 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     6121 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     6181 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     6241 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     6301 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     6361 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     6421 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     6481 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     6541 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     6601 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     6661 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     6721 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     6781 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     6841 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     6901 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     6961 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     7021 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     7081 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     7141 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     7201 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     7261 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     7321 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     7381 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     7441 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     7501 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     7561 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     7621 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     7681 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     7741 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7801 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7861 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     7921 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7981 gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     8041 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     8101 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     8161 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     8221 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     8281 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     8341 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     8401 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     8461 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     8521 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     8581 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     8641 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     8701 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     8761 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     8821 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     8881 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     8941 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     9001 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     9061 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     9121 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     9181 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     9241 cgatccccta tggt
//
