LOCUS       V038315                 7948 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038315
VERSION     V038315
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..3438
                     /label="Ntrk3(NM_008746)"
                     /note="Ntrk3(NM_008746)"
                     /gene="Ntrk3"
     CDS             3445..3510
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3548..3772
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3818..4246
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4260..4589
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4656..5450
                     /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    5624..5745
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5794..5810)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5818..5834
                     /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(5842..5872)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5887..5908
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6196..6781)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6952..7812)
                     /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(7813..7917)
                     /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 accatggatg tctctctttg cccagccaag tgtagtttct ggcggatttt cttgctggga
     1021 agcgtctggc tggactatgt gggctccgtg ctggcttgcc ctgcaaattg tgtctgcagc
     1081 aagactgaga tcaattgccg gcggccggac gatgggaacc tcttccctct cctggaaggg
     1141 caggattcag ggaacagcaa tgggaacgcc agcatcaaca ttacggacat ctcaaggaat
     1201 atcacttcca tacacataga gaactggcga ggcctgcaca cactcaatgc tgtggacatg
     1261 gagctctaca cgggactcca gaagctgacc atcaagaatt ctggactccg gaacatccag
     1321 cccagagcct tcgccaagaa cccccacttg cgttatataa acttgtcaag taaccggctc
     1381 accacactct cctggcagct cttccagacg ctgagccttc gggaattgag actggagcag
     1441 aacttcttca actgcagctg tgacatccgc tggatgcagc tgtggcagga acagggggag
     1501 gcgcggctgg acagccagag cctttactgc atcagtgctg atggctccca actccccctc
     1561 ttccgcatga acatcagtca gtgtgatctc ccagagatca gtgtgagcca cgtcaacctg
     1621 actgtccgag aaggagacaa tgccgtgatc acttgcaatg gctctggctc tcctttgcct
     1681 gatgtggact ggatagtcac tgggctgcag tccatcaaca cccaccagac caatctgaac
     1741 tggaccaatg tacatgccat caacttgacc ctggtgaacg tgacgagcga ggacaatggc
     1801 ttcaccctga cgtgcattgc agagaacgtg gtgggcatga gcaatgccag tgttgctctc
     1861 actgtctact accctccacg tgtggtgagc ctggtggagc ctgaggtacg cctggaacat
     1921 tgcattgagt ttgtggtgcg tggcaacccg acacccacgc tccactggct gtacaatgga
     1981 cagccattga gggagtccaa gatcattcac atggactact accaggaggg ggaggtctca
     2041 gagggctgcc tgctcttcaa caagcccacc cactacaaca atggcaacta caccctcatt
     2101 gctaagaatg ccctgggcac ggccaaccag accatcaacg gccacttcct gaaggagccc
     2161 tttccagaga gcacagattt ctttgacttt gagtctgatg cgagccctac acctcctatc
     2221 actgtgaccc acaaaccaga ggaagacact tttggggtgt ccatagcagt cggacttgct
     2281 gcctttgcct gcgtccttct ggtggttctc tttatcatga tcaacaagta tggtcgccgg
     2341 tccaaatttg gaatgaaggg tcctgtggct gttatcagtg gagaggagga ctcagccagc
     2401 ccactgcatc acatcaacca tggcatcact acaccatcat cgttggatgc tgggcctgac
     2461 acagtggtca ttggcatgac ccgcatccca gtcattgaga acccccagta cttccgtcag
     2521 ggtcacaatt gccacaagcc agacacatat gttcagcaca tcaagaggag agacatcgtg
     2581 ttgaagagag aattgggtga gggagccttt gggaaggtct tcctggctga gtgctacaat
     2641 ctaagcccca ccaaagacaa gatgctagtg gcagtgaagg ccctgaagga tcccaccttg
     2701 gctgccagga aggatttcca gagggaggct gagctgctca cgaacctgca gcatgagcat
     2761 attgtcaagt tctatggggt gtgtggtgat ggtgacccac tcatcatggt ctttgaatac
     2821 atgaagcatg gagaccttaa caagttcctc agggcccatg ggccagatgc catgatcctc
     2881 gtggatggac agccacgtca ggccaagggg gagctagggc tctctcagat gctccacatc
     2941 gccagtcaga tagcctcggg catggtgtac ctggcttccc agcactttgt acaccgggac
     3001 ctggccacga ggaactgcct ggttggagcc aatctactag tgaagattgg agattttggc
     3061 atgtccaggg acgtctacag tactgattac tacagggtgg gaggacacac catgctcccc
     3121 atccgctgga tgccccctga aagcataatg taccggaagt tcaccacaga gagtgatgtc
     3181 tggagcttcg gggttattct ttgggagatc tttacctatg ggaagcaacc atggttccag
     3241 ctttccaaca cggaggtcat tgaatgcatc acccaaggcc gtgtcttgga gagacccaga
     3301 gtctgcccta aagaagtgta tgatgtcatg ctggggtgct ggcagaggga accacagcag
     3361 cggctgaata ttaaggagat ctacaaaatc ctccatgctt tggggaaggc caccccgatc
     3421 tacctggaca ttcttggcct cgaggactac aaagaccatg acggtgatta taaagatcat
     3481 gacatcgact acaaggatga cgatgacaag tagtgagggc ccgtttaaac ccgctgatca
     3541 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3601 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3661 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3721 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     3781 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     3841 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3901 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3961 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4021 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4081 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4141 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4201 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     4261 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4321 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4381 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4441 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4501 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     4561 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     4621 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4681 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4741 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4801 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     4861 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4921 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4981 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5041 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5101 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5161 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     5221 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5281 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5341 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5401 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5461 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5521 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5581 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     5641 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5701 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     5761 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5821 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5881 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5941 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6001 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6061 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6121 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6181 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6241 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6301 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6361 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6421 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6481 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6541 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6601 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     6661 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6721 cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6781 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6841 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6901 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6961 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7021 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7081 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7141 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7201 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7261 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7321 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7381 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7441 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7501 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7561 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7621 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7681 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7741 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7801 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7861 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7921 atttccccga aaagtgccac ctgacgtc
//
