LOCUS       V019672                 7196 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019672
VERSION     V019672
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..1876
                     /label="TACR2(NM_001057)"
                     /note="TACR2(NM_001057)"
                     /gene="TACR2"
     CDS             1883..1948
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1972..2560
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2626..2674
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2876..3304
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3318..3647
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3714..4508
                     /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    4684..4805
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4854..4870)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4878..4894
                     /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(4902..4932)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4947..4968
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5256..5844)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6015..6875)
                     /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(6876..6980)
                     /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 ccatggggac ctgtgacatt gtgactgaag ccaatatctc
      721 atctggccct gagagcaaca ccacgggcat cacagccttc tccatgccca gctggcaact
      781 ggcactgtgg gccacagcct acctggccct ggtgctggtg gccgtgacgg gtaatgccat
      841 cgtcatctgg atcatcctgg cccatcggag gatgcgcaca gtcaccaact acttcatcgt
      901 caatctggcg ctggctgacc tctgcatggc tgccttcaat gccgccttca actttgtcta
      961 tgccagccac aacatctggt actttggccg tgccttctgc tacttccaga acctcttccc
     1021 catcacagcc atgtttgtca gcatctactc catgaccgcc attgctgccg acaggtacat
     1081 ggccatcgtc caccccttcc agcctcggct ttcagctccc agcaccaagg cggttattgc
     1141 tggcatctgg ctggtggctc tcgccctggc ctcccctcag tgcttctact ccaccgtcac
     1201 catggaccag ggtgccacca agtgcgtggt ggcctggccc gaagacagcg ggggcaagac
     1261 gctcctcctg taccacctcg tggtgatcgc cctcatctac ttcctgccgc tcgcggtgat
     1321 gtttgtagcc tacagcgtca tcggcctcac gctctggagg cgcgcagtgc ccggacatca
     1381 ggcgcacggt gccaacctgc gccacctgca ggccatgaag aagtttgtga agaccatggt
     1441 gctggtggtg ctgacgtttg ccatctgctg gctgccctac cacctctact tcatcctggg
     1501 cagcttccag gaggacatct actgccacaa gttcatccag caagtctacc tggcactctt
     1561 ctggttggcc atgagctcta ccatgtacaa tcccatcatc tactgctgtc tcaaccacag
     1621 gtttcgctct ggattccggc ttgccttccg ctgctgccca tgggtcacac ccaccaagga
     1681 agataagctc gagctgactc ccacgacctc cctctccacg agagtcaaca ggtgtcacac
     1741 taaggagact ttgttcatgg ctggggacac agccccctcc gaggctacca gtggggaggc
     1801 ggggcgtccc caggatggat cagggctatg gtttgggtat ggtttgcttg cccccaccaa
     1861 aactcatgtt gaaattctcg aggactacaa agaccatgac ggtgattata aagatcatga
     1921 catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga caatcaacct
     1981 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     2041 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     2101 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     2161 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     2221 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     2281 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     2341 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     2401 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     2461 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     2521 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa
     2581 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     2641 agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     2701 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     2761 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     2821 gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg
     2881 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2941 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3001 gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct
     3061 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3121 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3181 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3241 attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3301 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3361 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3421 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3481 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3541 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     3601 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     3661 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     3721 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3781 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3841 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     3901 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     3961 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4021 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4081 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4141 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4201 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     4261 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4321 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4381 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4441 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4501 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca
     4561 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4621 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4681 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4741 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4801 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     4861 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4921 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4981 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5041 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5101 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5161 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5221 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5281 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5341 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5401 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc
     5461 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5521 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5581 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5641 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     5701 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5761 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5821 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5881 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5941 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6001 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6061 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6121 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6181 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6241 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6301 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6361 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6421 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6481 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6541 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6601 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6661 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6721 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6781 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6841 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6901 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6961 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     7021 ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt
     7081 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     7141 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg
//
