LOCUS       V018714                10154 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018714
VERSION     V018714
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..5644
                     /label="PLXNC1(NM_005761)"
                     /note="PLXNC1(NM_005761)"
                     /gene="PLXNC1"
     CDS             5651..5716
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    5754..5978
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      6024..6452
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6466..6795
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6862..7656
                     /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    7830..7951
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(8000..8016)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8024..8040
                     /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(8048..8078)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8093..8114
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8402..8987)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9158..10018)
                     /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(10019..10123)
                     /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 atggaggtct cccggaggaa
      961 ggcgccgccg cgccccccgc gccccgcagc gccactgccc ctgctcgcct atctgctggc
     1021 actggcggct cccggccggg gcgcggacga gcccgtgtgg cggtcggagc aagccatcgg
     1081 agccatcgcg gcgagccagg aggacggcgt gtttgtggcg agcggcagct gcctggacca
     1141 gctggactac agcctggagc acagcctctc gcgcctgtac cgggaccaag cgggcaactg
     1201 cacagagccg gtctcgctgg cgccccccgc gcggccccgg cccgggagca gcttcagcaa
     1261 gctgctgctg ccctaccgcg agggggcggc cggcctcggg gggctgctgc tcaccggctg
     1321 gaccttcgac cggggcgcct gcgaggtgcg gcccctgggc aacctgagcc gcaactccct
     1381 gcgcaacggc accgaggtgg tgtcgtgcca cccgcagggc tcgacggccg gcgtggtgta
     1441 ccgcgcgggc cggaacaacc gctggtacct ggcggtggcc gccacctacg tgctgcctga
     1501 gccggagacg gcgagccgct gcaaccccgc ggcatccgac cacgacacgg ccatcgcgct
     1561 caaggacacg gaggggcgca gcctggccac gcaggagctg gggcgcctca agctgtgcga
     1621 gggcgcgggc agcctgcact tcgtggacgc ctttctctgg aacggcagca tctacttccc
     1681 ctactacccc tacaactaca cgagcggcgc tgccaccggc tggcccagca tggcgcgcat
     1741 cgcgcagagc accgaggtgc tgttccaggg ccaggcatcc ctcgactgcg gccacggcca
     1801 ccccgacggc cgccgcctgc tcctctcctc cagcctagtg gaggccctgg acgtctgggc
     1861 gggagtgttc agcgcggccg ctggagaggg ccaggagcgg cgctccccca ccaccacggc
     1921 gctctgcctc ttcagaatga gtgagatcca ggcgcgcgcc aagagggtca gctgggactt
     1981 caagacggcc gagagccact gcaaagaagg ggatcaacct gaaagagtcc aaccaatcgc
     2041 atcatctacc ttgatccatt ccgacctgac atccgtttat ggcaccgtgg taatgaacag
     2101 gactgtttta ttcttgggga ctggagatgg ccagttactt aaggttattc ttggtgagaa
     2161 tttgacttca aattgtccag aggttatcta tgaaattaaa gaagagacac ctgttttcta
     2221 caaactcgtt cctgatcctg tgaagaatat ctacatttat ctaacagctg ggaaagaggt
     2281 gaggagaatt cgtgttgcaa actgcaataa acataaatcc tgttcggagt gtttaacagc
     2341 cacagaccct cactgcggtt ggtgccattc gctacaaagg tgcacttttc aaggagattg
     2401 tgtacattca gagaacttag aaaactggct ggatatttcg tctggagcaa aaaagtgccc
     2461 taaaattcag ataattcgaa gcagtaaaga aaagactaca gtgactatgg tgggaagctt
     2521 ctctccaaga cactcaaagt gcatggtgaa gaatgtggac tctagcaggg agctctgcca
     2581 gaataaaagt cagcccaacc ggacctgcac ctgtagcatc ccaaccagag caacctacaa
     2641 agatgtttca gttgtcaacg tgatgttctc cttcggttct tggaatttat cagacagatt
     2701 caactttacc aactgctcat cattaaaaga atgcccagca tgcgtagaaa ctggctgcgc
     2761 gtggtgtaaa agtgcaagaa ggtgtatcca ccccttcaca gcttgcgacc cttctgatta
     2821 tgagagaaac caggaacagt gtccagtggc tgtcgagaag acatcaggag gaggaagacc
     2881 caaggagaac aaggggaaca gaaccaacca ggctttacag gtcttctaca ttaagtccat
     2941 tgagccacag aaagtatcga cattagggaa aagcaacgtg atagtaacgg gagcaaactt
     3001 tacccgggca tcgaacatca caatgatcct gaaaggaacc agtacctgtg ataaggatgt
     3061 gatacaggtt agccatgtgc taaatgacac ccacatgaaa ttctctcttc catcaagccg
     3121 gaaagaaatg aaggatgtgt gtatccagtt tgatggtggg aactgctctt ctgtgggatc
     3181 cttatcctac attgctctgc cacattgttc ccttatattt cctgctacca cctggatcag
     3241 tggtggtcaa aatataacca tgatgggcag aaattttgat gtaattgaca acttaatcat
     3301 ttcacatgaa ttaaaaggaa acataaatgt ctctgaatat tgtgtggcga cttactgcgg
     3361 gtttttagcc cccagtttaa agagttcaaa agtgcgcacg aatgtcactg tgaagctgag
     3421 agtacaagac acctacttgg attgtggaac cctgcagtat cgggaggacc ccagattcac
     3481 ggggtatcgg gtggaatccg aggtggacac agaactggaa gtgaaaattc aaaaagaaaa
     3541 tgacaacttc aacatttcca aaaaagacat tgaaattact ctcttccatg gggaaaatgg
     3601 gcaattaaat tgcagttttg aaaatattac tagaaatcaa gatcttacca ccatcctttg
     3661 caaaattaaa ggcatcaaga ctgcaagcac cattgccaac tcttctaaga aagttcgggt
     3721 caagctggga aacctggagc tctacgtcga gcaggagtca gttccttcca catggtattt
     3781 tctgattgtg ctccctgtct tgctagtgat tgtcattttt gcggccgtgg gggtgaccag
     3841 gcacaaatcg aaggagctga gtcgcaaaca gagtcaacaa ctagaattgc tggaaagcga
     3901 gctccggaaa gagatacgtg acggctttgc tgagctgcag atggataaat tggatgtggt
     3961 tgatagtttt ggaactgttc ccttccttga ctacaaacat tttgctctga gaactttctt
     4021 ccctgagtca ggtggcttca cccacatctt cactgaagat atgcataaca gagacgccaa
     4081 cgacaagaat gaaagtctca cagctttgga tgccctaatc tgtaataaaa gctttcttgt
     4141 tactgtcatc cacacccttg aaaagcagaa gaacttttct gtgaaggaca ggtgtctgtt
     4201 tgcctccttc ctaaccattg cactgcaaac caagctggtc tacctgacca gcatcctaga
     4261 ggtgctgacc agggacttga tggaacagtg tagtaacatg cagccgaaac tcatgctgag
     4321 acgcacggag tccgtcgtcg aaaaactcct cacaaactgg atgtccgtct gcctttctgg
     4381 atttctccgg gagactgtcg gagagccctt ctatttgctg gtgacgactc tgaaccagaa
     4441 aattaacaag ggtcccgtgg atgtaatcac ttgcaaagcc ctgtacacac ttaatgaaga
     4501 ctggctgttg tggcaggttc cggaattcag tactgtggca ttaaacgtcg tctttgaaaa
     4561 aatcccggaa aacgagagtg cagatgtctg tcggaatatt tcagtcaatg ttctcgactg
     4621 tgacaccatt ggccaagcca aagaaaagat tttccaagca ttcttaagca aaaatggctc
     4681 tccttatgga cttcagctta atgaaattgg tcttgagctt caaatgggca cacgacagaa
     4741 agaacttctg gacatcgaca gttcctccgt gattcttgaa gatggaatca ccaagctaaa
     4801 caccattggc cactatgaga tatcaaatgg atccactata aaagtcttta agaagatagc
     4861 aaattttact tcagatgtgg agtactcgga tgaccactgc catttgattt taccagattc
     4921 ggaagcattc caagatgtgc aaggaaagag acatcgaggg aagcacaagt tcaaagtaaa
     4981 agaaatgtat ctgacaaagc tgctgtcgac caaggtggca attcattctg tgcttgaaaa
     5041 actttttaga agcatttgga gtttacccaa cagcagagct ccatttgcta taaaatactt
     5101 ttttgacttt ttggacgccc aggctgaaaa caaaaaaatc acagatcctg acgtcgtaca
     5161 tatttggaaa acaaacagcc ttcctcttcg cttctgggta aacatcctga agaaccctca
     5221 gtttgtcttt gacattaaga agacaccaca tatagacggc tgtttgtcag tgattgccca
     5281 ggcattcatg gatgcatttt ctctcacaga gcagcaacta gggaaggaag caccaactaa
     5341 taagcttctc tatgccaagg atatcccaac ctacaaagaa gaagtaaaat cttattacaa
     5401 agcaatcagg gatttgcctc cattgtcatc ctcagaaatg gaagaatttt taactcagga
     5461 atctaagaaa catgaaaatg aatttaatga agaagtggcc ttgacagaaa tttacaaata
     5521 catcgtaaaa tattttgatg agattctaaa taaactagaa agagaacgag ggctggaaga
     5581 agctcagaaa caactcttgc atgtaaaagt cttatttgat gaaaagaaga aatgcaagtg
     5641 gatgctcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa
     5701 ggatgacgat gacaagtagt gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     5761 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     5821 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     5881 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     5941 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     6001 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     6061 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     6121 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     6181 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     6241 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     6301 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     6361 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     6421 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     6481 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     6541 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     6601 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     6661 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     6721 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     6781 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     6841 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     6901 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     6961 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     7021 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     7081 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     7141 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     7201 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     7261 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     7321 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     7381 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     7441 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     7501 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     7561 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     7621 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     7681 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     7741 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     7801 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     7861 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     7921 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     7981 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     8041 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     8101 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     8161 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     8221 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     8281 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     8341 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     8401 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     8461 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     8521 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     8581 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     8641 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     8701 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     8761 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     8821 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     8881 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt
     8941 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     9001 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     9061 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     9121 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     9181 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     9241 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     9301 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     9361 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     9421 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     9481 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     9541 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     9601 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     9661 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     9721 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     9781 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     9841 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     9901 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     9961 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
    10021 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
    10081 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
    10141 tgccacctga cgtc
//
