LOCUS       V040931                 6718 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040931
VERSION     V040931
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             964..2208
                     /label="PTPN2(NM_002828)"
                     /note="PTPN2(NM_002828)"
                     /gene="PTPN2"
     CDS             2215..2280
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2318..2542
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2588..3016
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3030..3359
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3426..4220
                     /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    4394..4515
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4564..4580)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4588..4604
                     /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(4612..4642)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4657..4678
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4966..5551)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5722..6582)
                     /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(6583..6687)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatgccca ccaccatcga gcgggagttc gaagagttgg atactcagcg tcgctggcag
     1021 ccgctgtact tggaaattcg aaatgagtcc catgactatc ctcatagagt ggccaagttt
     1081 ccagaaaaca gaaatcgaaa cagatacaga gatgtaagcc catatgatca cagtcgtgtt
     1141 aaactgcaaa atgctgagaa tgattatatt aatgccagtt tagttgacat agaagaggca
     1201 caaaggagtt acatcttaac acagggtcca cttcctaaca catgctgcca tttctggctt
     1261 atggtttggc agcagaagac caaagcagtt gtcatgctga accgcattgt ggagaaagaa
     1321 tcggttaaat gtgcacagta ctggccaaca gatgaccaag agatgctgtt taaagaaaca
     1381 ggattcagtg tgaagctctt gtcagaagat gtgaagtcgt attatacagt acatctacta
     1441 caattagaaa atatcaatag tggtgaaacc agaacaatat ctcactttca ttatactacc
     1501 tggccagatt ttggagtccc tgaatcacca gcttcatttc tcaatttctt gtttaaagtg
     1561 agagaatctg gctccttgaa ccctgaccat gggcctgcgg tgatccactg tagtgcaggc
     1621 attgggcgct ctggcacctt ctctctggta gacacttgtc ttgttttgat ggaaaaagga
     1681 gatgatatta acataaaaca agtgttactg aacatgagaa aataccgaat gggtcttatt
     1741 cagaccccag atcaactgag attctcatac atggctataa tagaaggagc aaaatgtata
     1801 aagggagatt ctagtataca gaaacgatgg aaagaacttt ctaaggaaga cttatctcct
     1861 gcctttgatc attcaccaaa caaaataatg actgaaaaat acaatgggaa cagaataggt
     1921 ctagaagaag aaaaactgac aggtgaccga tgtacaggac tttcctctaa aatgcaagat
     1981 acaatggagg agaacagtga gagtgctcta cggaaacgta ttcgagagga cagaaaggcc
     2041 accacagctc agaaggtgca gcagatgaaa cagaggctaa atgagaatga acgaaaaaga
     2101 aaaaggtggt tatattggca acctattctc actaagatgg ggtttatgtc agtcattttg
     2161 gttggcgctt ttgttggctg gacactgttt tttcagcaaa atgccctact cgaggactac
     2221 aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag
     2281 tagtgagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     2341 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     2401 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     2461 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     2521 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     2581 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2641 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     2701 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     2761 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     2821 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     2881 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     2941 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3001 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3061 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3121 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3181 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3241 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     3301 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     3361 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     3421 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3481 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3541 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     3601 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     3661 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3721 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     3781 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     3841 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     3901 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     3961 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4021 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4081 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4141 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4201 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     4261 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4321 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4381 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4441 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4501 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     4561 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4621 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4681 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4741 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4801 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4861 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4921 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4981 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5041 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5101 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5161 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5221 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5281 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5341 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5401 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5461 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca
     5521 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5581 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5641 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5701 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5761 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5821 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5881 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5941 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6001 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6061 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6121 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6181 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6241 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6301 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6361 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6421 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6481 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6541 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6601 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6661 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
