LOCUS       V041921                 6886 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041921
VERSION     V041921
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             928..2322
                     /label="TACR3(NM_001059)"
                     /note="TACR3(NM_001059)"
                     /gene="TACR3"
     CDS             2329..2352
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             2359..2448
                     /codon_start=1
                     /product="two Strep-Tag®II moieties connected by a linker
                     for enhanced binding to Strep-Tactin®,&nbsp;an engineered
                     form of streptavidin (Schmidt et al., 2013)"
                     /transl_table=1
                     /translation="SA,WSHPQFEK,GGGSGGGSGGSA,WSHPQFEK"
                     /label="Twin-Strep-tag"
     polyA_signal    2486..2710
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2756..3184
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3198..3527
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3594..4388
                     /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    4562..4683
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4732..4748)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4756..4772
                     /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(4780..4810)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4825..4846
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5134..5719)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5890..6750)
                     /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(6751..6855)
                     /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 aagctgaatt cgccaccatg gccactctcc cagcagcaga aacctggata
      961 gacgggggtg gaggcgtggg tgcagacgcc gtgaacctga ccgcctcgct agctgccggg
     1021 gcggccacgg gggcagttga gactgggtgg ctgcaactgc tggaccaagc tggcaacctc
     1081 tcctcctccc cttccgcgct gggactgcct gtggcttccc ccgcgccctc ccagccctgg
     1141 gccaacctca ccaaccagtt cgtgcagccg tcctggcgca tcgcgctctg gtccctggcg
     1201 tatggtgtgg tggtggcagt ggcagttttg ggaaatctca tcgtcatctg gatcatcctg
     1261 gcccacaagc gcatgaggac tgtcaccaac tacttccttg tgaacctggc tttctccgac
     1321 gcctccatgg ccgccttcaa cacgttggtc aatttcatct acgcgcttca tagcgagtgg
     1381 tactttggcg ccaactactg ccgcttccag aacttctttc ctatcacagc tgtgttcgcc
     1441 agcatctact ccatgacggc cattgcggtg gacaggtata tggctattat tgatcccttg
     1501 aaacccagac tgtctgctac agcaaccaag attgtcattg gaagtatttg gattctagca
     1561 tttctacttg ccttccctca gtgtctttat tccaaaacca aagtcatgcc aggccgtact
     1621 ctctgctttg tgcaatggcc agaaggtccc aaacaacatt tcacttacca tattatcgtc
     1681 attatactgg tgtactgttt cccattgctc atcatgggta ttacatacac cattgttgga
     1741 attactctct ggggaggaga aatcccagga gatacctgtg acaagtatca tgagcagcta
     1801 aaggccaaaa gaaaggttgt caaaatgatg attattgttg tcatgacatt tgctatctgc
     1861 tggctgccct atcatattta cttcattctc actgcaatct atcaacaact aaatagatgg
     1921 aaatacatcc agcaggtcta cctggctagc ttttggctgg caatgagctc aaccatgtac
     1981 aatcccatca tctactgctg tctgaataaa agatttcgag ctggcttcaa gagagcattt
     2041 cgctggtgtc ctttcatcaa agtttccagc tatgatgagc tagagctcaa gaccaccagg
     2101 tttcatccaa accggcaaag cagtatgtac accgtgacca gaatggagtc catgacagtc
     2161 gtgtttgacc ccaacgatgc agacaccacc aggtccagtc ggaagaaaag agcaacgcca
     2221 agagacccaa gtttcaatgg ctgctctcgc aggaattcca aatctgcctc cgccacttca
     2281 agtttcataa gctcacccta tacctctgtg gatgaatatt ctctcgagga ttacaaggat
     2341 gacgacgata agggatccag cgcctggtcc caccctcagt ttgagaaggg cggaggctct
     2401 ggcggcggaa gcggaggatc tgcttggagc cacccccagt tcgaaaagtg atgagggccc
     2461 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     2521 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     2581 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     2641 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     2701 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     2761 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2821 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2881 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     2941 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3001 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3061 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3121 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3181 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3241 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3301 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3361 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3421 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     3481 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     3541 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     3601 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3661 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3721 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     3781 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     3841 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     3901 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     3961 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4021 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4081 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     4141 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4201 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4261 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4321 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4381 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4441 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4501 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4561 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4621 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4681 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     4741 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4801 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4861 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4921 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4981 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5041 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5101 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5161 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5221 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5281 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5341 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5401 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5461 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5521 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5581 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5641 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     5701 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5761 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5821 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5881 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5941 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6001 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6061 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6121 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6181 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6241 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6301 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6361 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6421 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6481 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6541 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6601 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6661 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6721 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6781 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6841 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
