LOCUS       V022295                 6164 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022295
VERSION     V022295
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..1687
                     /label="TNFSF12(NM_003809)"
                     /note="TNFSF12(NM_003809)"
                     /gene="TNFSF12"
     CDS             1703..1726
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     polyA_signal    1764..1988
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2034..2462
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2476..2805
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             2872..3666
                     /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    3840..3961
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4010..4026)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4034..4050
                     /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(4058..4088)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4103..4124
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4412..4997)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5168..6028)
                     /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(6029..6133)
                     /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 atggccgccc gtcggagcca
      961 gaggcggagg gggcgccggg gggagccggg caccgccctg ctggtcccgc tcgcgctggg
     1021 cctgggcctg gcgctggcct gcctcggcct cctgctggcc gtggtcagtt tggggagccg
     1081 ggcatcgctg tccgcccagg agcctgccca ggaggagctg gtggcagagg aggaccagga
     1141 cccgtcggaa ctgaatcccc agacagaaga aagccaggat cctgcgcctt tcctgaaccg
     1201 actagttcgg cctcgcagaa gtgcacctaa aggccggaaa acacgggctc gaagagcgat
     1261 cgcagcccat tatgaagttc atccacgacc tggacaggac ggagcgcagg caggtgtgga
     1321 cgggacagtg agtggctggg aggaagccag aatcaacagc tccagccctc tgcgctacaa
     1381 ccgccagatc ggggagttta tagtcacccg ggctgggctc tactacctgt actgtcaggt
     1441 gcactttgat gaggggaagg ctgtctacct gaagctggac ttgctggtgg atggtgtgct
     1501 ggccctgcgc tgcctggagg aattctcagc cactgcggcg agttccctcg ggccccagct
     1561 ccgcctctgc caggtgtctg ggctgttggc cctgcggcca gggtcctccc tgcggatccg
     1621 caccctcccc tgggcccatc tcaaggctgc ccccttcctc acctacttcg gactcttcca
     1681 ggttcactgg cggccgctcg aggattacaa ggatgacgac gataagtagt gagggcccgt
     1741 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     1801 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     1861 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     1921 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     1981 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     2041 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2101 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2161 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2221 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     2281 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2341 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     2401 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     2461 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2521 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2581 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     2641 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     2701 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     2761 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     2821 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     2881 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     2941 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3001 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3061 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3121 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3181 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3241 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3301 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3361 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     3421 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3481 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     3541 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     3601 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     3661 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     3721 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     3781 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     3841 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3901 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     3961 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4021 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4081 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4141 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4201 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4261 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4321 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4381 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4441 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4501 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4561 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4621 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4681 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4741 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4801 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4861 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4921 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca
     4981 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5041 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5101 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5161 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5221 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5281 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5341 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5401 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5461 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5521 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5581 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5641 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5701 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5761 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5821 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5881 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5941 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6001 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6061 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6121 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
