LOCUS       V037013                 8309 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037013
VERSION     V037013
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             908..3742
                     /label="UNC5B(NM_170744)"
                     /note="UNC5B(NM_170744)"
                     /gene="UNC5B"
     CDS             3743..3769
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3773..3799
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3803..3829
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    3876..4100
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4146..4574
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4588..4917
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4984..5778
                     /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    5952..6073
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6122..6138)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6146..6162
                     /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(6170..6200)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6215..6236
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6524..7109)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7280..8140)
                     /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(8141..8245)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg ggggcccgga gcggagctcg gggcgcgctg ctgctggcac tgctgctctg
      961 ctgggacccg aggctgagcc aagcaggcac tgattctggc agcgaggtgc tccctgactc
     1021 cttcccgtca gcgccagcag agccgctgcc ctacttcctg caggagccac aggacgccta
     1081 cattgtgaag aacaagcctg tggagctccg ctgccgcgcc ttccccgcca cacagatcta
     1141 cttcaagtgc aacggcgagt gggtcagcca gaacgaccac gtcacacagg aaggcctgga
     1201 tgaggccacc ggcctgcggg tgcgcgaggt gcagatcgag gtgtcgcggc agcaggtgga
     1261 ggagctcttt gggctggagg attactggtg ccagtgcgtg gcctggagct ccgcgggcac
     1321 caccaagagt cgccgagcct acgtccgcat cgcctacctg cgcaagaact tcgatcagga
     1381 gcctctgggc aaggaggtgc ccctggacca tgaggttctc ctgcagtgcc gcccgccgga
     1441 gggggtgcct gtggccgagg tggaatggct caagaatgag gatgtcatcg accccaccca
     1501 ggacaccaac ttcctgctca ccatcgacca caacctcatc atccgccagg cccgcctgtc
     1561 ggacactgcc aactatacct gcgtggccaa gaacatcgtg gccaaacgcc ggagcaccac
     1621 tgccaccgtc atcgtctacg tgaatggcgg ctggtccagc tgggcagagt ggtcaccctg
     1681 ctccaaccgc tgtggccgag gctggcagaa gcgcacccgg acctgcacca accccgctcc
     1741 actcaacgga ggggccttct gcgagggcca ggcattccag aagaccgcct gcaccaccat
     1801 ctgcccagtc gatggggcgt ggacggagtg gagcaagtgg tcagcctgca gcactgagtg
     1861 tgcccactgg cgtagccgcg agtgcatggc gcccccaccc cagaacggag gccgtgactg
     1921 cagcgggacg ctgctcgact ctaagaactg cacagatggg ctgtgcatgc aaaataagaa
     1981 aactctaagc gaccccaaca gccacctgct ggaggcctca ggggatgcgg cgctgtatgc
     2041 ggggctcgtg gtggccatct tcgtggtcgt ggcaatcctc atggcggtgg gggtggtggt
     2101 gtaccgccgc aactgccgtg acttcgacac agacatcact gactcatctg ctgccctgac
     2161 tggtggtttc caccccgtca actttaagac ggcaaggccc agcaacccgc agctcctaca
     2221 cccctctgtg cctcctgacc tgacagccag cgccggcatc taccgcggac ccgtgtatgc
     2281 cctgcaggac tccaccgaca aaatccccat gaccaactct cctctgctgg accccttacc
     2341 cagccttaag gtcaaggtct acagctccag caccacgggc tctgggccag gcctggcaga
     2401 tggggctgac ctgctggggg tcttgccgcc tggcacatac cctagcgatt tcgcccggga
     2461 cacccacttc ctgcacctgc gcagcgccag cctcggttcc cagcagctct tgggcctgcc
     2521 ccgagaccca gggagcagcg tcagcggcac ctttggctgc ctgggtggga ggctcagcat
     2581 ccccggcaca ggggtcagct tgctggtgcc caatggagcc attccccagg gcaagttcta
     2641 cgagatgtat ctactcatca acaaggcaga aagtaccctc ccgctttcag aagggaccca
     2701 gacagtattg agcccctcgg tgacctgtgg acccacaggc ctcctgctgt gccgccccgt
     2761 catcctcacc atgccccact gtgccgaagt cagtgcccgt gactggatct ttcagctcaa
     2821 gacccaggcc caccagggcc actgggagga ggtggtgacc ctggatgagg agaccctgaa
     2881 cacaccctgc tactgccagc tggagcccag ggcctgtcac atcctgctgg accagctggg
     2941 cacctacgtg ttcacgggcg agtcctattc ccgctcagca gtcaagcggc tccagctggc
     3001 cgtcttcgcc cccgccctct gcacctccct ggagtacagc ctccgggtct actgcctgga
     3061 ggacacgcct gtagcactga aggaggtgct ggagctggag cggactctgg gcggatactt
     3121 ggtggaggag ccgaaaccgc taatgttcaa ggacagttac cacaacctgc gcctctccct
     3181 ccatgacctc ccccatgccc attggaggag caagctgctg gccaaatacc aggagatccc
     3241 cttctatcac atttggagtg gcagccagaa ggccctccac tgcactttca ccctggagag
     3301 gcacagcttg gcctccacag agctcacctg caagatctgc gtgcggcaag tggaagggga
     3361 gggccagata ttccagctgc ataccactct ggcagagaca cctgctggct ccctggacac
     3421 tctctgctct gcccctggca gcactgtcac cacccagctg ggaccttatg ccttcaagat
     3481 cccactgtcc atccgccaga agatatgcaa cagcctagat gcccccaact cacggggcaa
     3541 tgactggcgg atgttagcac agaagctctc tatggaccgg tacctgaatt actttgccac
     3601 caaagcgagc cccacgggtg tgatcctgga cctctgggaa gctctgcagc aggacgatgg
     3661 ggacctcaac agcctggcga gtgccttgga ggagatgggc aagagtgaga tgctggtggc
     3721 tgtggccacc gacggggact gctacccata cgacgtacct gattatgcaa cttatcctta
     3781 tgatgtacct gattatgcta gctatcctta tgatgtccca gactacgcat agctcgagtc
     3841 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     3901 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     3961 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     4021 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     4081 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     4141 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4201 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4261 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4321 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4381 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4441 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4501 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4561 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4621 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4681 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4741 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4801 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4861 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4921 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     4981 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     5041 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     5101 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5161 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5221 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5281 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5341 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5401 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5461 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     5521 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5581 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5641 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5701 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5761 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5821 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5881 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5941 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6001 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6061 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     6121 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6181 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6241 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6301 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6361 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6421 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6481 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6541 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6601 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6661 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6721 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6781 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6841 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6901 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6961 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7021 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc
     7081 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7141 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7201 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7261 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7321 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7381 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7441 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7501 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7561 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7621 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7681 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7741 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7801 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7861 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7921 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7981 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8041 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8101 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8161 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8221 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     8281 gatcgggaga tctcccgatc ccctatggt
//
