LOCUS       V020813                 9098 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020813
VERSION     V020813
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..3778
                     /label="RTN3(NM_001265589)"
                     /note="RTN3(NM_001265589)"
                     /gene="RTN3"
     CDS             3785..3850
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3874..4462
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4528..4576
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4778..5206
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5220..5549
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5616..6410
                     /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    6586..6707
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6756..6772)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6780..6796
                     /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(6804..6834)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6849..6870
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7158..7746)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7917..8777)
                     /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(8778..8882)
                     /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 ccatggcgga gccgtcggcg gccactcagt cccattccat
      721 ctcctcgtcg tccttcggag ccgagccgtc cgcgcccggc ggcggcggga gcccaggagc
      781 ctgccccgcc ctggggacga agagctgcag ctcctcctgt gcggattcct ttgtttcttc
      841 ctcttcctct cagcctgtat ctctattttc gacctcacaa gagggattga gctctctttg
      901 ctctgatgag ccatcttcag aaattatgac ttcttccttt ctttcatctt ctgaaataca
      961 taacactggc cttacaatac tacatggaga aaaaagccat gtgttaggga gccagcctat
     1021 tttagccaaa gaaggaaaag accacttgga tcttctagat atgaaaaaga tggaaaagcc
     1081 tcaggggacc agcaacaacg tatcagactc ttcagtttct cttgcagcag gagttcattg
     1141 tgaccgtcct tctattccag ccagtttccc agagcatcct gcttttctct caaagaaaat
     1201 tggtcaagtg gaagagcaaa tagataaaga gaccaagaac ccaaatgggg tatcaagtag
     1261 ggaggctaaa actgcattgg atgctgatga cagattcact ttgctgacag cccagaaacc
     1321 acctactgag tactctaagg tagaaggcat ttatacatat tctttgtctc catccaaagt
     1381 ttcaggagat gatgttattg aaaaggattc ccctgaatca ccatttgaag taattattga
     1441 caaagcagca tttgacaaag aatttaaaga ctcatataag gagagcacag atgattttgg
     1501 tagctggtct gtgcacactg ataaagaatc atccgaagac atttcagaga ctaatgacaa
     1561 gctttttcca ctgagaaata aagaggcagg acgttaccca atgtctgcat tgctcagtag
     1621 gcagttttca cacacaaatg cagcactgga agaggtgtcc agatgcgtga atgatatgca
     1681 taactttact aacgaaatac tgacttggga tctggttccc caagtgaaac aacagaccga
     1741 taaatcttct gactgcatca caaaaactac aggacttgac atgagtgaat ataattcaga
     1801 aattccagtt gtaaatctta aaactagcac tcatcagaaa actcctgtat gttctattga
     1861 tgggagcact cccatcacta aatcaacagg tgattgggca gaagcatctc tccagcaaga
     1921 aaatgctatt actggaaaac ctgtacctga ctctttgaat tccacaaaag aattcagtat
     1981 caaaggtgtg caaggcaata tgcagaaaca ggatgacaca cttgcagaat tacctggatc
     2041 tccacctgag aaatgtgact ctttgggttc tggagtggcc acagtgaaag tggttttacc
     2101 tgatgaccac ctgaaagatg aaatggactg gcagagctct gcattgggag aaatcacaga
     2161 agctgatagt tctggtgagt ctgatgacac agtaatagag gacatcacag cagatacatc
     2221 atttgaaaat aacaaaattc aggctgaaaa acctgtttcc attccaagtg ctgttgtaaa
     2281 aacaggtgaa agagaaatca aagagattcc cagttgtgag agagaagaaa aaacatctaa
     2341 aaactttgaa gaattggtca gtgactctga gctgcatcaa gatcagcctg atattcttgg
     2401 aaggagtcca gctagtgagg cagcatgttc aaaagtaccc gatacgaatg tctccttaga
     2461 agatgtgagt gaagttgctc ctgaaaagcc tattactact gagaacccca aacttccttc
     2521 aacagtgtct ccaaatgttt ttaatgagac agaattctca ttaaatgtga caacatctgc
     2581 ctatttggag tcattacatg ggaaaaatgt taaacatata gatgattcct ccccagagga
     2641 cctgatagca gcctttacag aaaccagaga taaaggaata gtagatagtg aaagaaatgc
     2701 ttttaaagca atatcagaga agatgacaga ctttaaaaca actcctcctg tagaagtctt
     2761 acatgaaaat gagtccggtg gttctgaaat taaagacatt ggaagcaaat acagtgaaca
     2821 aagcaaagaa acaaatggaa gtgagcctct aggtgttttc cctacccaag gtactccagt
     2881 agcatctctt gacttagaac aagaacagct cacaattaag gctcttaaag aattaggtga
     2941 aagacaggtt gagaagtcaa cttctgcaca gcgtgacgca gaattgcctt ctgaagaagt
     3001 actgaagcaa actttcacat ttgctccaga atcttggcca cagagatcat atgacatcct
     3061 agaacgtaat gtcaagaatg gatctgatct tgggatttcc cagaagccca tcactatcag
     3121 agaaactact agggtagatg ctgtttccag ccttagcaag actgaattgg taaaaaagca
     3181 tgtcctagca agacttctga cagacttctc agtgcacgat ctgattttct ggagagatgt
     3241 gaagaagact gggtttgtct ttggcaccac gctgatcatg ctgctttccc tggcagcttt
     3301 cagtgtcatc agtgtggttt cttacctcat cctggctctt ctctctgtca ccatcagctt
     3361 caggatctac aagtccgtca tccaagctgt acagaagtca gaagaaggcc atccattcaa
     3421 agcctacctg gacgtagaca ttactctgtc ctcagaagct ttccataatt acatgaatgc
     3481 tgccatggtg cacatcaaca gggccctgaa actcattatt cgtctctttc tggtagaaga
     3541 tctggttgac tccttgaagc tggctgtctt catgtggctg atgacctatg ttggtgctgt
     3601 ttttaacgga atcacccttc taattcttgc tgaactgctc attttcagtg tcccgattgt
     3661 ctatgagaag tacaagaccc agattgatca ctatgttggc atcgcccgag atcagaccaa
     3721 gtcaattgtt gaaaagatcc aagcaaaact ccctggaatc gccaaaaaaa aggcagaact
     3781 cgaggactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga
     3841 cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta caaaatttgt
     3901 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     3961 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     4021 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     4081 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     4141 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     4201 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     4261 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     4321 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     4381 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     4441 tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca cggaaggaga
     4501 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt
     4561 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     4621 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     4681 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     4741 gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     4801 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4861 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4921 gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4981 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     5041 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     5101 acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc
     5161 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     5221 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     5281 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     5341 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     5401 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     5461 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     5521 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     5581 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     5641 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     5701 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     5761 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     5821 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     5881 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     5941 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     6001 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     6061 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     6121 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     6181 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     6241 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     6301 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     6361 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     6421 ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     6481 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     6541 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     6601 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     6661 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     6721 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     6781 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6841 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6901 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6961 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     7021 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     7081 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     7141 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     7201 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     7261 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     7321 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     7381 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7441 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7501 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7561 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     7621 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7681 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7741 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7801 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7861 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7921 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7981 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     8041 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     8101 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     8161 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     8221 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     8281 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     8341 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     8401 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     8461 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     8521 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     8581 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     8641 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     8701 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     8761 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8821 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8881 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     8941 tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     9001 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     9061 aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
