LOCUS       V022794                 7802 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022794
VERSION     V022794
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             683..2458
                     /label="TGFBR2(NM_001024847)"
                     /note="TGFBR2(NM_001024847)"
                     /gene="TGFBR2"
     CDS             2465..2554
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    2578..3166
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3232..3280
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3482..3910
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3924..4253
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4320..5114
                     /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    5290..5411
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5460..5476)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5484..5500
                     /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(5508..5538)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5553..5574
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5862..6450)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6621..7481)
                     /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(7482..7586)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatgggtcg ggggctgctc aggggcctgt ggccgctgca
      721 catcgtcctg tggacgcgta tcgccagcac gatcccaccg cacgttcaga agtcggatgt
      781 ggaaatggag gcccagaaag atgaaatcat ctgccccagc tgtaatagga ctgcccatcc
      841 actgagacat attaataacg acatgatagt cactgacaac aacggtgcag tcaagtttcc
      901 acaactgtgt aaattttgtg atgtgagatt ttccacctgt gacaaccaga aatcctgcat
      961 gagcaactgc agcatcacct ccatctgtga gaagccacag gaagtctgtg tggctgtatg
     1021 gagaaagaat gacgagaaca taacactaga gacagtttgc catgacccca agctccccta
     1081 ccatgacttt attctggaag atgctgcttc tccaaagtgc attatgaagg aaaaaaaaaa
     1141 gcctggtgag actttcttca tgtgttcctg tagctctgat gagtgcaatg acaacatcat
     1201 cttctcagaa gaatataaca ccagcaatcc tgacttgttg ctagtcatat ttcaagtgac
     1261 aggcatcagc ctcctgccac cactgggagt tgccatatct gtcatcatca tcttctactg
     1321 ctaccgcgtt aaccggcagc agaagctgag ttcaacctgg gaaaccggca agacgcggaa
     1381 gctcatggag ttcagcgagc actgtgccat catcctggaa gatgaccgct ctgacatcag
     1441 ctccacgtgt gccaacaaca tcaaccacaa cacagagctg ctgcccattg agctggacac
     1501 cctggtgggg aaaggtcgct ttgctgaggt ctataaggcc aagctgaagc agaacacttc
     1561 agagcagttt gagacagtgg cagtcaagat ctttccctat gaggagtatg cctcttggaa
     1621 gacagagaag gacatcttct cagacatcaa tctgaagcat gagaacatac tccagttcct
     1681 gacggctgag gagcggaaga cggagttggg gaaacaatac tggctgatca ccgccttcca
     1741 cgccaagggc aacctacagg agtacctgac gcggcatgtc atcagctggg aggacctgcg
     1801 caagctgggc agctccctcg cccgggggat tgctcacctc cacagtgatc acactccatg
     1861 tgggaggccc aagatgccca tcgtgcacag ggacctcaag agctccaata tcctcgtgaa
     1921 gaacgaccta acctgctgcc tgtgtgactt tgggctttcc ctgcgtctgg accctactct
     1981 gtctgtggat gacctggcta acagtgggca ggtgggaact gcaagataca tggctccaga
     2041 agtcctagaa tccaggatga atttggagaa tgttgagtcc ttcaagcaga ccgatgtcta
     2101 ctccatggct ctggtgctct gggaaatgac atctcgctgt aatgcagtgg gagaagtaaa
     2161 agattatgag cctccatttg gttccaaggt gcgggagcac ccctgtgtcg aaagcatgaa
     2221 ggacaacgtg ttgagagatc gagggcgacc agaaattccc agcttctggc tcaaccacca
     2281 gggcatccag atggtgtgtg agacgttgac tgagtgctgg gaccacgacc cagaggcccg
     2341 tctcacagcc cagtgtgtgg cagaacgctt cagtgagctg gagcatctgg acaggctctc
     2401 ggggaggagc tgctcggagg agaagattcc tgaagacggc tccctaaaca ctaccaaact
     2461 cgaggaacaa aaactaatat cagaggaaga tttggaacag aaactaatca gtgaagaaga
     2521 tttagagcaa aagttaattt ccgaggagga cttatagtga gggcccgata tctcgacaat
     2581 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     2641 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     2701 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     2761 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     2821 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     2881 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     2941 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     3001 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     3061 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     3121 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg aaacggggga
     3181 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     3241 aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     3301 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     3361 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     3421 gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc
     3481 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3541 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3601 acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt
     3661 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3721 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3781 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3841 taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3901 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3961 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4021 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4081 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4141 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4201 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4261 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     4321 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4381 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4441 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4501 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4561 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4621 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4681 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4741 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4801 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     4861 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4921 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4981 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5041 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5101 acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac
     5161 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5221 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5281 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5341 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5401 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     5461 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5521 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5581 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5641 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5701 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5761 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5821 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5881 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5941 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6001 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     6061 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6121 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6181 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6241 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6301 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6361 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6421 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6481 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6541 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6601 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6661 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6721 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6781 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6841 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6901 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6961 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7021 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7081 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7141 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7201 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7261 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7321 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7381 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7441 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7501 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7561 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     7621 ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc tctgatgccg
     7681 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     7741 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     7801 gg
//
