LOCUS       pT3-EF1a-Il33(mouse)    6245 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043671
VERSION     V043671
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        1..1179
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
                     /label="EF-1α promoter"
     promoter        1196..1214
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     protein_bind    1293..1317
                     /bound_moiety="BP Clonase™"
                     /gene="mutant version of&nbsp;attB"
                     /note="recombination site for the Gateway® BP reaction"
                     /label="attB1"
     CDS             1348..2145
                     /label="Il33(NM_001164724)"
                     /note="Il33(NM_001164724)"
                     /gene="Il33"
     polyA_signal    2173..2397
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     protein_bind    2467..2500
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
                     /label="loxP"
     primer_bind     complement(2958..2974)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     promoter        3448..3552
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             3553..4413
                     /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"
     rep_origin      4584..5172
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    5460..5481
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        5496..5526
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5534..5550
                     /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"
     primer_bind     5558..5574
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6072..6105
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
                     /label="loxP"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtactaata
     1201 cgactcacta tagggagacc caagctggct aggtaagctt ggtaccgagc tcggatccac
     1261 tagtccagtg tggtggaatt ctgcagatat caacaagttt gtacaaaaaa gcaggctccg
     1321 cggccgcccc cttcaccatg ggccaccatg agacctagaa tgaagtattc caactccaag
     1381 atttccccgg caaagttcag cagcaccgca ggcgaagccc tggtcccgcc ttgcaaaata
     1441 agaagatccc aacagaagac caaagaattc tgccatgtct actgcatgag actccgttct
     1501 ggcctcacca taagaaagga gactagttat tttaggaaag aacccacgaa aagatattca
     1561 ctaaaatcgg gtaccaagca tgaagagaac ttctctgcct atccacggga ttctaggaag
     1621 agatccttgc ttggcagtat ccaagcattt gctgcgtctg ttgacacatt gagcatccaa
     1681 ggaacttcac ttttaacaca gtctcctgcc tccctgagta catacaatga ccaatctgtt
     1741 agttttgttt tggagaatgg atgttatgtg atcaatgttg acgactctgg aaaagaccaa
     1801 gagcaagacc aggtgctact acgctactat gagtctccct gtcctgcaag tcaatcaggc
     1861 gacggtgtgg atgggaagaa gctgatggtg aacatgagtc ccatcaaaga cacagacatc
     1921 tggctgcatg ccaacgacaa ggactactcc gtggagcttc aaaggggtga cgtctcgcct
     1981 ccggaacagg ccttcttcgt ccttcacaaa aagtcctcgg actttgtttc atttgaatgc
     2041 aagaatcttc ctggcactta cataggagta aaagataacc agctggctct agtggaggag
     2101 aaagatgaga gctgcaacaa tattatgttt aagctctcga aaatctaatt taaacccgct
     2161 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2221 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2281 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2341 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     2401 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     2461 cattaaataa cttcgtataa tgtatgctat acgaagttat atcgtatgca tgctttgcat
     2521 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
     2581 aattccttga aatacatcca caggtacacc tccaattgac tcaaatgatg tcaattagtc
     2641 tatcagaagc ttctaaagcc atgacatcat tttctggaat tttccaagct gtttaaaggc
     2701 acagtcaact tagtgtatgt aaacttctga cccactggaa ttgtgataca gtgaattata
     2761 agtgaaataa tctgtctgta aacaattgtt ggaaaaatga cttgtgtcat gcacaaagta
     2821 gatgtcctaa ctgacttgcc aaaactattg tttgttaaca agaaatttgt ggagtagttg
     2881 aaaaacgagt tttaatgact ccaacttaag tgtatgtaaa cttccgactt caactgtagg
     2941 gtaccgagct cgaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     3001 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     3061 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg
     3121 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     3181 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     3241 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     3301 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     3361 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     3421 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3481 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3541 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     3601 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     3661 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     3721 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     3781 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     3841 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     3901 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     3961 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     4021 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     4081 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     4141 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     4201 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     4261 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     4321 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     4381 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     4441 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4501 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4561 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4621 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4681 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4741 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4801 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4861 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4921 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4981 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5041 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5101 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5161 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5221 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     5281 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     5341 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     5401 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     5461 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     5521 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     5581 tacgccaagc ttgcatgcct gcaggtcgac tctagaccta tacagttgaa gtcggaagtt
     5641 tacatacact taagttggag tcattaaaac tcgtttttca actactccac aaatttcttg
     5701 ttaacaaaca atagttttgg caagtcagtt aggacatcta ctttgtgcat gacacaagtc
     5761 atttttccaa caattgttta cagacagatt atttcactta taattcactg tatcacaatt
     5821 ccagtgggtc agaagtttac atacactaag ttgactgtgc ctttaaacag cttggaaaat
     5881 tccagaaaat gatgtcatgg ctttagaagc ttctgataga ctaattgaca tcatttgagt
     5941 caattggagg tgtacctgtg gatgtatttc aaggggatcc agacatgata agatacattg
     6001 atgagtttgg acaaaccaca actagaatgc agcggccgct agctcgagat cttaattaaa
     6061 ctagtatcga tataacttcg tataatgtat gctatacgaa gttatatgta cgggccagat
     6121 atacgcgttg acattgatta ttgactaggc ttttgcaaaa agctttgcaa agatggataa
     6181 agttttaaac agagaggaat ctttgcagct aatggacctt ctaggtcttg aaaggagtgg
     6241 gaatt
//
