LOCUS       V019056                 8315 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019056
VERSION     V019056
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..2995
                     /label="SEMA3A(NM_006080)"
                     /note="SEMA3A(NM_006080)"
                     /gene="SEMA3A"
     CDS             3002..3067
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3091..3679
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3745..3793
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3995..4423
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4437..4766
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4833..5627
                     /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    5803..5924
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5973..5989)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5997..6013
                     /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(6021..6051)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6066..6087
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6375..6963)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7134..7994)
                     /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(7995..8099)
                     /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 ccatgggctg gttaactagg attgtctgtc ttttctgggg
      721 agtattactt acagcaagag caaactatca gaatgggaag aacaatgtgc caaggctgaa
      781 attatcctac aaagaaatgt tggaatccaa caatgtgatc actttcaatg gcttggccaa
      841 cagctccagt tatcatacct tccttttgga tgaggaacgg agtaggctgt atgttggagc
      901 aaaggatcac atattttcat tcgacctggt taatatcaag gattttcaaa agattgtgtg
      961 gccagtatct tacaccagaa gagatgaatg caagtgggct ggaaaagaca tcctgaaaga
     1021 atgtgctaat ttcatcaagg tacttaaggc atataatcag actcacttgt acgcctgtgg
     1081 aacgggggct tttcatccaa tttgcaccta cattgaaatt ggacatcatc ctgaggacaa
     1141 tatttttaag ctggagaact cacattttga aaacggccgt gggaagagtc catatgaccc
     1201 taagctgctg acagcatccc ttttaataga tggagaatta tactctggaa ctgcagctga
     1261 ttttatgggg cgagactttg ctatcttccg aactcttggg caccaccacc caatcaggac
     1321 agagcagcat gattccaggt ggctcaatga tccaaagttc attagtgccc acctcatctc
     1381 agagagtgac aatcctgaag atgacaaagt atactttttc ttccgtgaaa atgcaataga
     1441 tggagaacac tctggaaaag ctactcacgc tagaataggt cagatatgca agaatgactt
     1501 tggagggcac agaagtctgg tgaataaatg gacaacattc ctcaaagctc gtctgatttg
     1561 ctcagtgcca ggtccaaatg gcattgacac tcattttgat gaactgcagg atgtattcct
     1621 aatgaacttt aaagatccta aaaatccagt tgtatatgga gtgtttacga cttccagtaa
     1681 cattttcaag ggatcagccg tgtgtatgta tagcatgagt gatgtgagaa gggtgttcct
     1741 tggtccatat gcccacaggg atggacccaa ctatcaatgg gtgccttatc aaggaagagt
     1801 cccctatcca cggccaggaa cttgtcccag caaaacattt ggtggttttg actctacaaa
     1861 ggaccttcct gatgatgtta taacctttgc aagaagtcat ccagccatgt acaatccagt
     1921 gtttcctatg aacaatcgcc caatagtgat caaaacggat gtaaattatc aatttacaca
     1981 aattgtcgta gaccgagtgg atgcagaaga tggacagtat gatgttatgt ttatcggaac
     2041 agatgttggg accgttctta aagtagtttc aattcctaag gagacttggt atgatttaga
     2101 agaggttctg ctggaagaaa tgacagtttt tcgggaaccg actgctattt cagcaatgga
     2161 gctttccact aagcagcaac aactatatat tggttcaacg gctggggttg cccagctccc
     2221 tttacaccgg tgtgatattt acgggaaagc gtgtgctgag tgttgcctcg cccgagaccc
     2281 ttactgtgct tgggatggtt ctgcatgttc tcgctatttt cccactgcaa agagacgcac
     2341 aagacgacaa gatataagaa atggagaccc actgactcac tgttcagact tacaccatga
     2401 taatcaccat ggccacagcc ctgaagagag aatcatctat ggtgtagaga atagtagcac
     2461 atttttggaa tgcagtccga agtcgcagag agcgctggtc tattggcaat tccagaggcg
     2521 aaatgaagag cgaaaagaag agatcagagt ggatgatcat atcatcagga cagatcaagg
     2581 ccttctgcta cgtagtctac aacagaagga ttcaggcaat tacctctgcc atgcggtgga
     2641 acatgggttc atacaaactc ttcttaaggt aaccctggaa gtcattgaca cagagcattt
     2701 ggaagaactt cttcataaag atgatgatgg agatggctct aagaccaaag aaatgtccaa
     2761 tagcatgaca cctagccaga aggtctggta cagagacttc atgcagctca tcaaccaccc
     2821 caatctcaac acaatggatg agttctgtga acaagtttgg aaaagggacc gaaaacaacg
     2881 tcggcaaagg ccaggacata ccccagggaa cagtaacaaa tggaagcact tacaagaaaa
     2941 taagaaaggt agaaacagga ggacccacga atttgagagg gcacccagga gtgtcctcga
     3001 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga
     3061 tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa aatttgtgaa
     3121 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     3181 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     3241 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     3301 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     3361 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     3421 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     3481 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     3541 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     3601 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     3661 ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg aaggagacaa
     3721 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt
     3781 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     3841 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     3901 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca
     3961 gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     4021 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4081 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4141 ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4201 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4261 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4321 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat
     4381 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     4441 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4501 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4561 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4621 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4681 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     4741 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     4801 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4861 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4921 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4981 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     5041 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5101 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5161 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5221 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5281 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5341 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5401 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5461 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5521 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5581 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5641 gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5701 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5761 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     5821 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5881 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     5941 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6001 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6061 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6121 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     6181 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6241 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6301 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6361 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6421 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6481 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6541 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     6601 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6661 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6721 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6781 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     6841 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6901 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6961 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7021 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7081 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7141 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7201 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7261 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7321 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7381 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7441 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7501 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7561 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7621 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7681 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7741 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7801 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7861 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7921 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7981 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8041 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8101 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     8161 ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     8221 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     8281 gcttgaccga caattgcatg aagaatctgc ttagg
//
