LOCUS       V036506                 8223 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036506
VERSION     V036506
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2903
                     /label="Il31ra(NM_139299)"
                     /note="Il31ra(NM_139299)"
                     /gene="Il31ra"
     CDS             2910..2975
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2999..3587
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3653..3701
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3903..4331
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4345..4674
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4741..5535
                     /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    5711..5832
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5881..5897)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5905..5921
                     /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(5929..5959)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5974..5995
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6283..6871)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7042..7902)
                     /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(7903..8007)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgtg gaccttggca ctgtgggcat
      781 tctctttcct ctgcaaattc agcctggcag tcctgccgac taagccagag aacatttcct
      841 gcgtctttta cttcgacaga aatctgactt gcacttggag accagagaag gaaaccaatg
      901 ataccagcta cattgtgact ttgacttact cctatggaaa aagcaattat agtgacaatg
      961 ctacagaggc ttcatattct tttccccgtt cctgtgcaat gcccccagac atctgcagtg
     1021 ttgaagtaca agctcaaaat ggagatggta aagttaaatc tgacatcaca tattggcatt
     1081 taatctccat agcaaaaacc gaaccaccta taattttaag tgtgaatcca atttgtaata
     1141 gaatgttcca gatacaatgg aaaccgcgtg aaaagactcg tgggtttcct ttagtatgca
     1201 tgcttcggtt cagaactgtc aacagtagcc actggacgga agtcaatttt gaaaactgta
     1261 aacaggtctg caacctcaca ggacttcagg ctttcacaga atatgtcctg gctctacgat
     1321 tcaggttcaa tgactcaaga tattggagca agtggagcaa agaagaaacc agagtgacta
     1381 tggaggaagt tccacatgtc ctggacctgt ggagaattct ggaaccagca gacatgaacg
     1441 gagacaggaa ggtgcgattg ttgtggaaga aggcaagagg agcccccgtc ttggagaaaa
     1501 catttggcta ccacatacag tactttgcag agaacagcac taacctcaca gagataaaca
     1561 acatcaccac ccagcagtat gaactgcttc tgatgagcca ggcacactct gtgtccgtga
     1621 cttcttttaa ttctcttggc aagtcccaag aggccatcct gaggatccca gatgtccatg
     1681 agaagacctt ccagtacatt aagagcatga aggcctacat agccgagccc ctgttggtgg
     1741 tgaactggca aagctccatt cctgcggtgg acacttggat agtggagtgg ctcccagaag
     1801 ctgccatgtc gaagttccct gccctttcct gggaatctgt gtctcaggtc acgaactgga
     1861 ccatcgagca agataaacta aaacctttca catgctataa tatatcagtg tatccagtgt
     1921 tgggacaccg agttggagag ccgtattcaa tccaagctta tgccaaagaa ggaactccat
     1981 taaaaggtcc tgagaccagg gtggagaaca tcggtctgag gacagccacg atcacatgga
     2041 aggagattcc taagagtgct aggaatggat ttatcaacaa ttacactgta ttttaccaag
     2101 ctgaaggtgg aaaagaactc tccaagactg ttaactctca tgccctgcag tgtgacctgg
     2161 agtctctgac acgaaggacc tcttatactg tttgggtcat ggccagcacc agagctggag
     2221 gtaccaacgg ggtgagaata aacttcaaga cattgtcaat cagtgtgttt gaaattgtcc
     2281 ttctaacatc tctagttgga ggaggccttc ttctacttag catcaaaaca gtgacttttg
     2341 gcctcagaaa gccaaaccgg ttgactcccc tgtgttgtcc tgatgttccc aaccctgctg
     2401 aaagtagttt agccacatgg ctcggagatg gtttcaagaa gtcaaatatg aaggagactg
     2461 gaaactctgg ggacacagaa gacgtggtcc taaaaccatg tcccgtcccc gcggatctca
     2521 ttgacaagct ggtagtgaac tttgagaatt ttctggaagt agttttgaca gaggaagctg
     2581 gaaagggtca ggcgagcatt ttgggaggag aagcgaatga gtatgtgacc tccccgtcta
     2641 ggcccgacgg tcccccaggg aaaagtttta aagagccctc cgttttaact gaggttgctt
     2701 ctgaagactc ccacagcacg tgttccagaa tggcggacga ggcgtactca gaattggcca
     2761 ggcagccttc gtcttcctgt cagagtccag ggctatcgcc tccccgtgaa gaccaagctc
     2821 agaatccata tttgaaaaat tcagtgacaa ccagggaatt tcttgtgcat gagaatatcc
     2881 cagagcacag caaaggagaa gtcggatccg actacaaaga ccatgacggt gattataaag
     2941 atcatgacat cgactacaag gatgacgatg acaagtagtg agggcccgat atctcgacaa
     3001 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     3061 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     3121 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     3181 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     3241 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     3301 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     3361 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     3421 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     3481 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     3541 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg
     3601 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     3661 aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     3721 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     3781 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     3841 cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc
     3901 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3961 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4021 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt
     4081 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4141 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4201 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4261 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4321 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4381 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4441 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4501 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4561 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     4621 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     4681 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     4741 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4801 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4861 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4921 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4981 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5041 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5101 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5161 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5221 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     5281 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5341 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5401 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5461 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5521 gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa
     5581 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5641 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5701 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5761 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5821 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     5881 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5941 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6001 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6061 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6121 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6181 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6241 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6301 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6361 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6421 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     6481 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6541 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6601 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6661 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6721 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6781 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6841 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6901 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6961 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7021 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7081 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7141 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7201 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7261 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7321 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     7381 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7441 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7501 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7561 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7621 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7681 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7741 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7801 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7861 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7921 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7981 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     8041 cggatcggga gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc
     8101 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     8161 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     8221 agg
//
