LOCUS       V023168                 8261 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023168
VERSION     V023168
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             941..3751
                     /label="Ror1(NM_013845)"
                     /note="Ror1(NM_013845)"
                     /gene="Ror1"
     CDS             3758..3823
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3861..4085
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4131..4559
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4573..4902
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4969..5763
                     /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    5937..6058
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6107..6123)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6131..6147
                     /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(6155..6185)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6200..6221
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6509..7094)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7265..8125)
                     /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(8126..8230)
                     /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 atccgccacc atgcaccggc cgcgccgccg
      961 cgggacgcgc ccgccaccgc tggcgctgct ggccgcgctg ctgctggccg cacgcggggc
     1021 tgatgcccaa gaaacagagt tgtcagtcag tgctgagctg gtgcctacct cgtcctggaa
     1081 cacttcaagt gaaatcgaca aaggttctta cttaaccctt gatgagccga tgaataacat
     1141 cacaacgtcc ctggggcaga ctgcagaact gcactgcaaa gtgtctggga atccacctcc
     1201 cagtatccgc tggttcaaga atgatgcacc tgtggtccaa gaacctcgga gaatctcctt
     1261 ccgggcaacc aactatggct ctcggctgcg gattagaaac cttgacacca cagacactgg
     1321 ttacttccag tgtgtggcaa caaatggcaa gaaagtggtg tctaccactg gtgtcctgtt
     1381 tgtcaaattt gggcctcctc cgaccgcaag cccaggatcc tcagatgagt atgaagaaga
     1441 tggattctgt cagccgtacc gaggcattgc atgtgcacga tttattggca accgcactgt
     1501 gtatatggag tctttgcata tgcaggggga aatagaaaat cagatcacag ctgccttcac
     1561 catgattggc acctccagcc atttatctga taagtgctct cagttcgcca tcccttcctt
     1621 gtgccactac gccttcccgt actgtgacga aacctcatct gtcccaaagc cccgtgactt
     1681 gtgtcgtgat gaatgtgaag tgctggagaa tgtcctgtgt cagacagagt acatttttgc
     1741 cagatcaaat cccatgattt tgatgaggct gaagttacca aactgtgagg atctccccca
     1801 gccagagagc ccggaagctg caaactgcat acggattggc attcccatgg cggatcctat
     1861 aaataaaaat cacaaatgct acaatagcac gggtgtagac taccggggaa ccgtcagtgt
     1921 gaccaagtct ggacgccagt gccagccatg gaattctcag tacccacaca cacacagctt
     1981 cactgctctg cgctttccgg agctcaacgg aggccactcc tactgccgca accctggcaa
     2041 ccagaaggaa gctccctggt gctttacctt ggatgaaaac tttaagtctg acctgtgtga
     2101 catcccagca tgtgattcca aagattccaa agagaagaat aaaatggaaa tcttgtacat
     2161 tctggtgcca agtgtggcca ttcccctggc tatcgccttc ctcttcttct tcatctgtgt
     2221 gtgccgcaat aaccagaagt cttcatcacc accagtccag aggcagccaa aacccgtcag
     2281 aggacagaat gtggagatgt ccatgctcaa tgcatacaag cccaagagca aggctaaaga
     2341 gctgcctctt tccgctgtgc gtttcatgga agaattgggt gaatgtacct ttggaaaaat
     2401 ctataagggc catctctacc tcccaggcat ggaccatgct cagctggtgg ctatcaaaac
     2461 cttgaaagac tataacaacc cccagcagtg gacagaattt caacaggaag cctctctcat
     2521 ggctgaacta caccacccca atattgtatg cctcctcgga gccgtcaccc aggaacaacc
     2581 tgtgtgtatg ctctttgagt atatgaacca gggagacctc cacgagttcc tcatcatgcg
     2641 atccccgcat tccgatgtcg gctgtagcag tgatgaagat gggacggtca aatccagcct
     2701 ggaccatggt gatttcttac acatagcaat tcagatcgca gctggcatgg agtacctgtc
     2761 tagtcacttc ttcgtgcaca aggaccttgc agctcgcaac attttaattg gagagcaact
     2821 gcatgtaaaa atttcagatc ttgggctttc cagagaaatt tactctgctg attactatag
     2881 ggtacagagt aagtcttcac tgcccatccg ctggatgccc cctgaggcca tcatgtacgg
     2941 caaattctcc tccgattccg atatctggtc tttcggggtt gtgttgtggg agattttcag
     3001 ctttggactc cagccatatt atgggtttag taatcaggaa gtgattgaaa tggtgcggaa
     3061 gcggcagctc ttaccatgtt ctgaagactg cccgccgcgc atgtacagcc tcatgaccga
     3121 gtgctggaat gagatacctt ccaggagacc acgctttaaa gacatccacg tccggcttcg
     3181 atcctgggag ggcctctcaa gtcacaccag ctctaccacc ccctcgggtg gaaatgccac
     3241 cacgcagacc acttccctca gtgccagccc tgtgagtaac ctcagcaacc cccgatttcc
     3301 caattacatg ttcccgagcc aagggattac accccagggt cagatcgctg gtttcattgg
     3361 cccagcgata cctcagaacc agcgcttcat ccccatcaat ggatacccaa tacctcctgg
     3421 ctatgcagcc tttccagctg cccactacca gcctgcaggg cctcccaggg tgattcagca
     3481 ctgcccacct ccgaagagtc ggtccccaag cagcgccagc ggatcgacca gcactggcca
     3541 tgtggccagc ttgccctcat caggatccaa tcaggaagca aacgttcctt tgctacccca
     3601 catgtcaatt ccaaatcacc ctggtggaat gggtatcact gtttttggca acaaatctca
     3661 aaaaccgtac aaaatagact caaaacaatc gtctttgctt ggggactccc atatccatgg
     3721 gcacaccgaa tctatgattt ctgcagaagt gctcgaggac tacaaagacc atgacggtga
     3781 ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag ggcccgttta
     3841 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3901 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3961 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     4021 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     4081 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     4141 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4201 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4261 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4321 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4381 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4441 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4501 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     4561 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4621 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4681 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4741 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4801 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     4861 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     4921 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4981 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     5041 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     5101 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     5161 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     5221 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     5281 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     5341 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     5401 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     5461 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     5521 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5581 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5641 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5701 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5761 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     5821 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5881 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     5941 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     6001 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     6061 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     6121 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6181 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6241 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     6301 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6361 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6421 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6481 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6541 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6601 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6661 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6721 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6781 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6841 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6901 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6961 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7021 gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     7081 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     7141 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7201 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7261 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7321 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7381 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7441 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7501 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7561 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7621 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7681 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7741 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7801 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7861 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7921 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7981 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     8041 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     8101 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     8161 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8221 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
