LOCUS       V036728                 8354 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036728
VERSION     V036728
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..3073
                     /label="Stat3(NM_011486)"
                     /note="Stat3(NM_011486)"
                     /gene="Stat3"
     CDS             3110..3802
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="red fluorescent protein from Entacmaea
                     quadricolor"
                     /transl_table=1
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
                     /label="TurboRFP"
     CDS             3809..3874
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3921..4145
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4191..4619
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4633..4962
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5029..5823
                     /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    5997..6118
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6167..6183)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6191..6207
                     /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(6215..6245)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6260..6281
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6569..7154)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7325..8185)
                     /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(8186..8290)
                     /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 gctcagtgga accagctgca gcagctggac acacgctacc tggagcagct
      961 gcaccagctg tacagcgaca gcttccccat ggagctgcgg cagttcctgg caccttggat
     1021 tgagagtcaa gactgggcat atgcagccag caaagagtca catgccacgt tggtgtttca
     1081 taatctcttg ggtgaaattg accagcaata tagccgattc ctgcaagagt ccaatgtcct
     1141 ctatcagcac aaccttcgaa gaatcaagca gtttctgcag agcaggtatc ttgagaagcc
     1201 aatggaaatt gcccggatcg tggcccgatg cctgtgggaa gagtctcgcc tcctccagac
     1261 ggcagccacg gcagcccagc aagggggcca ggccaaccac ccaacagccg ccgtagtgac
     1321 agagaagcag cagatgttgg agcagcatct tcaggatgtc cggaagcgag tgcaggatct
     1381 agaacagaaa atgaaggtgg tggagaacct ccaggacgac tttgatttca actacaaaac
     1441 cctcaagagc caaggagaca tgcaggatct gaatggaaac aaccagtctg tgaccagaca
     1501 gaagatgcag cagctggaac agatgctcac agccctggac cagatgcgga gaagcattgt
     1561 gagtgagctg gcggggctct tgtcagcaat ggagtacgtg cagaagacac tgactgatga
     1621 agagctggct gactggaaga ggcggcagca gatcgcgtgc atcggaggcc ctcccaacat
     1681 ctgcctggac cgtctggaaa actggataac ttcattagca gaatctcaac ttcagacccg
     1741 ccaacaaatt aagaaactgg aggagctgca gcagaaagtg tcctacaagg gcgaccctat
     1801 cgtgcagcac cggcccatgc tggaggagag gatcgtggag ctgttcagaa acttaatgaa
     1861 gagtgccttc gtggtggagc ggcagccctg catgcccatg cacccggacc ggcccttagt
     1921 catcaagact ggtgtccagt ttaccacgaa agtcaggttg ctggtcaaat ttcctgagtt
     1981 gaattatcag cttaaaatta aagtgtgcat tgataaagac tctggggatg ttgctgccct
     2041 cagagggtct cggaaattta acattctggg cacgaacaca aaagtgatga acatggagga
     2101 gtctaacaac ggcagcctgt ctgcagagtt caagcacctg acccttaggg agcagagatg
     2161 tgggaatgga ggccgtgcca attgtgatgc ctccttgatc gtgactgagg agctgcacct
     2221 gatcaccttc gagactgagg tgtaccacca aggcctcaag attgacctag agacccactc
     2281 cttgccagtt gtggtgatct ccaacatctg tcagatgcca aatgcttggg catcaatcct
     2341 gtggtataac atgctgacca ataaccccaa gaacgtgaac ttcttcacta agccgccaat
     2401 tggaacctgg gaccaagtgg ccgaggtgct cagctggcag ttctcgtcca ccaccaagcg
     2461 ggggctgagc atcgagcagc tgacaacgct ggctgagaag ctcctagggc ctggtgtgaa
     2521 ctactcaggg tgtcagatca catgggctaa attctgcaaa gaaaacatgg ctggcaaggg
     2581 cttctccttc tgggtctggc tagacaatat catcgacctt gtgaaaaagt atatcttggc
     2641 cctttggaat gaagggtaca tcatgggttt catcagcaag gagcgggagc gggccatcct
     2701 aagcacaaag cccccgggca ccttcctact gcgcttcagc gagagcagca aagaaggagg
     2761 ggtcactttc acttgggtgg aaaaggacat cagtggcaag acccagatcc agtctgtaga
     2821 gccatacacc aagcagcagc tgaacaacat gtcatttgct gaaatcatca tgggctataa
     2881 gatcatggat gcgaccaaca tcctggtgtc tccacttgtc tacctctacc ccgacattcc
     2941 caaggaggag gcatttggaa agtactgtag gcccgagagc caggagcacc ccgaagccga
     3001 cccaggtagt gctgccccgt acctgaagac caagttcatc tgtgtgacac cattcattga
     3061 tgcagtttgg aaagaattcg gtggaggcgg atcaggaggc ggtggatcta tgagcgagct
     3121 gatcaaggag aacatgcaca tgaagctgta catggagggc accgtgaaca accaccactt
     3181 caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca tgaagatcaa
     3241 ggtggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacca gcttcatgta
     3301 cggcagcaaa gccttcatca accacaccca gggcatcccc gacttcttta agcagtcctt
     3361 ccctgagggc ttcacatggg agagaatcac cacatacgaa gacgggggcg tgctgaccgc
     3421 tacccaggac accagcttcc agaacggctg catcatctac aacgtcaaga tcaacggggt
     3481 gaacttccca tccaacggcc ctgtgatgca gaagaaaaca cgcggctggg aggccaacac
     3541 cgagatgctg taccccgctg acggcggcct gagaggccac agccagatgg ccctgaagct
     3601 cgtgggcggg ggctacctgc actgctcctt caagaccaca tacagatcca agaaacccgc
     3661 taagaacctc aagatgcccg gcttccactt cgtggaccac agactggaaa gaatcaagga
     3721 ggccgacaaa gagacctacg tcgagcagca cgagatggct gtggccaagt actgcgacct
     3781 ccctagcaaa ctggggcaca gactcgagga ctacaaagac catgacggtg attataaaga
     3841 tcatgacatc gactacaagg atgacgatga caagtagctc gagtctagag ggcccgttta
     3901 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3961 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     4021 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     4081 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     4141 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     4201 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4261 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4321 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4381 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4441 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4501 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4561 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     4621 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4681 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4741 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4801 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4861 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     4921 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     4981 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     5041 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     5101 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     5161 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     5221 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     5281 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     5341 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     5401 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     5461 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     5521 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     5581 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5641 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5701 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5761 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5821 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     5881 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5941 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     6001 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     6061 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     6121 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     6181 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6241 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6301 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     6361 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6421 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6481 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6541 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6601 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6661 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6721 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6781 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6841 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6901 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6961 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     7021 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7081 gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     7141 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     7201 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7261 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7321 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7381 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7441 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7501 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7561 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7621 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7681 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7741 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7801 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7861 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7921 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7981 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     8041 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     8101 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     8161 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     8221 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8281 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     8341 cgatccccta tggt
//
