LOCUS       V026712                 8036 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026712
VERSION     V026712
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..3526
                     /label="IL12RB2(NM_001559)"
                     /note="IL12RB2(NM_001559)"
                     /gene="IL12RB2"
     CDS             3533..3598
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3636..3860
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3906..4334
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4348..4677
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4744..5538
                     /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    5712..5833
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5882..5898)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5906..5922
                     /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(5930..5960)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5975..5996
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6284..6869)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7040..7900)
                     /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(7901..8005)
                     /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 atggcacata cttttagagg
      961 atgctcattg gcatttatgt ttataatcac gtggctgttg attaaagcaa aaatagatgc
     1021 gtgcaagaga ggcgatgtga ctgtgaagcc ttcccatgta attttacttg gatccactgt
     1081 caatattaca tgctctttga agcccagaca aggctgcttt cactattcca gacgtaacaa
     1141 gttaatcctg tacaagtttg acagaagaat caattttcac catggccact ccctcaattc
     1201 tcaagtcaca ggtcttcccc ttggtacaac cttgtttgtc tgcaaactgg cctgtatcaa
     1261 tagtgatgaa attcaaatat gtggagcaga gatcttcgtt ggtgttgctc cagaacagcc
     1321 tcaaaattta tcctgcatac agaagggaga acaggggact gtggcctgca cctgggaaag
     1381 aggacgagac acccacttat acactgagta tactctacag ctaagtggac caaaaaattt
     1441 aacctggcag aagcaatgta aagacattta ttgtgactat ttggactttg gaatcaacct
     1501 cacccctgaa tcacctgaat ccaatttcac agccaaggtt actgctgtca atagtcttgg
     1561 aagctcctct tcacttccat ccacattcac attcttggac atagtgaggc ctcttcctcc
     1621 gtgggacatt agaatcaaat ttcaaaaggc ttctgtgagc agatgtaccc tttattggag
     1681 agatgaggga ctggtactgc ttaatcgact cagatatcgg cccagtaaca gcaggctctg
     1741 gaatatggtt aatgttacaa aggccaaagg aagacatgat ttgctggatc tgaaaccatt
     1801 tacagaatat gaatttcaga tttcctctaa gctacatctt tataagggaa gttggagtga
     1861 ttggagtgaa tcattgagag cacaaacacc agaagaagag cctactggga tgttagatgt
     1921 ctggtacatg aaacggcaca ttgactacag tagacaacag atttctcttt tctggaagaa
     1981 tctgagtgtc tcagaggcaa gaggaaaaat tctccactat caggtgacct tgcaggagct
     2041 gacaggaggg aaagccatga cacagaacat cacaggacac acctcctgga ccacagtcat
     2101 tcctagaacc ggaaattggg ctgtggctgt gtctgcagca aattcaaaag gcagttctct
     2161 gcccactcgt attaacataa tgaacctgtg tgaggcaggg ttgctggctc ctcgccaggt
     2221 ctctgcaaac tcagagggca tggacaacat tctggtgact tggcagcctc ccaggaaaga
     2281 tccctctgct gttcaggagt acgtggtgga atggagagag ctccatccag ggggtgacac
     2341 acaggtccct ctaaactggc tacggagtcg accctacaat gtgtctgctc tgatttcaga
     2401 gaacataaaa tcctacatct gttatgaaat ccgtgtgtat gcactctcag gggatcaagg
     2461 aggatgcagc tccatcctgg gtaactctaa gcacaaagca ccactgagtg gcccccacat
     2521 taatgccatc acagaggaaa aggggagcat tttaatttca tggaacagca ttccagtcca
     2581 ggagcaaatg ggctgcctcc tccattatag gatatactgg aaggaacggg actccaactc
     2641 ccagcctcag ctctgtgaaa ttccctacag agtctcccaa aattcacatc caataaacag
     2701 cctgcagccc cgagtgacat atgtcctgtg gatgacagct ctgacagctg ctggtgaaag
     2761 ttcccacgga aatgagaggg aattttgtct gcaaggtaaa gccaattgga tggcgtttgt
     2821 ggcaccaagc atttgcattg ctatcatcat ggtgggcatt ttctcaacgc attacttcca
     2881 gcaaaaggtg tttgttctcc tagcagccct cagacctcag tggtgtagca gagaaattcc
     2941 agatccagca aatagcactt gcgctaagaa atatcccatt gcagaggaga agacacagct
     3001 gcccttggac aggctcctga tagactggcc cacgcctgaa gatcctgaac cgctggtcat
     3061 cagtgaagtc cttcatcaag tgaccccagt tttcagacat cccccctgct ccaactggcc
     3121 acaaagggaa aaaggaatcc aaggtcatca ggcctctgag aaagacatga tgcacagtgc
     3181 ctcaagccca ccacctccaa gagctctcca agctgagagc agacaactgg tggatctgta
     3241 caaggtgctg gagagcaggg gctccgaccc aaagcccgaa aacccagcct gtccctggac
     3301 ggtgctccca gcaggtgacc ttcccaccca tgatggctac ttaccctcca acatagatga
     3361 cctcccctca catgaggcac ctctcgctga ctctctggaa gaactggagc ctcagcacat
     3421 ctccctttct gttttcccct caagttctct tcacccactc accttctcct gtggtgataa
     3481 gctgactctg gatcagttaa agatgaggtg tgactccctc atgctcctcg aggactacaa
     3541 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta
     3601 gtgagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     3661 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     3721 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     3781 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     3841 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     3901 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3961 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4021 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4081 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4141 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4201 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4261 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4321 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4381 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4441 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4501 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4561 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4621 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4681 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     4741 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4801 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4861 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4921 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4981 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5041 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5101 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5161 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5221 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     5281 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5341 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5401 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5461 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5521 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5581 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5641 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5701 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5761 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5821 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     5881 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5941 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6001 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6061 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6121 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6181 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6241 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6301 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6361 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6421 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6481 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6541 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6601 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6661 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6721 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6781 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc
     6841 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6901 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6961 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7021 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7081 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7141 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7201 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7261 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7321 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7381 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7441 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7501 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7561 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7621 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7681 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7741 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7801 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7861 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7921 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7981 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
