LOCUS       pSG66:_Shaobin_version_of_pOR21PR-UTR1-KcsAKv1.3-sfGFP-6His-T500        3809 bp    DNA     circular UNA 20-SEP-2017
DEFINITION  prep from 031416 is 100% correct. gigaprep is also correct(one aa
            in linker is different from template).
ACCESSION   urn.local.shaobinguo.3s-824h23q
VERSION     urn.local.shaobinguo.3s-824h23q
KEYWORDS    .
SOURCE      
  ORGANISM  
            .
FEATURES             Location/Qualifiers
     source          order(1..>38,<1317..3809)
                     /dnas_title="pBEST-OR2-OR1-Pr-UTR1-deGFP-T500"
                     /label="source"
     PCR_product     join(3721..3809,<1317..1386,1..>38)
                     /pair="Pair 1"
                     /set="800bp_amp"
                     /current=1
                     /score="0.000"
                     /modified_by="User"
                     /label="868bp-linear fragment"
     PCR_product     join(3497..3809,<1317..1613,1..>38)
                     /pair="Pair 3"
                     /set="1200bp_amp"
                     /current=0
                     /score="0.264"
                     /modified_by="User"
                     /label="1319bp-linear fragment"
     PCR_product     join(3253..3809,<1317..1840,1..>38)
                     /pair="Pair 1"
                     /set="1600bp_amp"
                     /current=1
                     /score="1.000"
                     /modified_by="User"
                     /label="1768bp-linear fragment"
     misc_feature    1..33
                     /note="UTR1"
                     /label="UTR1"
     ligation        35..38
                     /label="Ligation"
     source          <35..>530
                     /clone_id=3884304
                     /organism="Homo sapiens"
                     /requester="rfachini"
                     /clone="pAMG21-MDG-coKcsA/KcsA(Chi IV) #-C6"
                     /vntifkey=98
                     /label="source Homo sapiens"
     misc_feature    35
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     CDS             36..>530
                     /vntifkey=84
                     /modified_by="User"
                     /label="KcsA-Kv1.3"
     misc_feature    <39..>530
                     /Source="referenceToDocument: urn:local:.:3x-824h256"
                     /label="Digestion of ZS5061 piece 2: Kscav1.3 extraction
                     by BsaI - Fragment 2 extraction (concatenated sequence 2)"
                     /note="Geneious type: Concatenated sequence"
     primer_bind     164..181
                     /Sequence="CGTACTGCTTGCTGGTTC"
                     /Hairpin_Tm="None"
                     /Tm="56.1"
                     /Self_Dimer_Tm="None"
                     /%GC="55.6"
                     /modified_by="User"
                     /label="Oligo_81f"
     misc_feature    263
                     /Original_Bases="G"
                     /label="G"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    complement(333..349)
                     /Sequence="ATGCCAGCCACCATAAC"
                     /Hairpin_Tm="None"
                     /Tm="54.3"
                     /Self_Dimer_Tm="None"
                     /%GC="52.9"
                     /modified_by="User"
                     /label="Oligo_81r"
                     /note="Geneious type: primer_bind_reverse"
     misc_difference 531..584
                     /created_by="User"
                     /label="Linker"
     misc_feature    <531..557
                     /Source="referenceToDocument: urn:local:.:3y-824h256"
                     /label="TetR (3) pcr (concatenated sequence 2)"
                     /note="Geneious type: Concatenated sequence"
     misc_feature    531..1298
                     /Original_Bases="TAAGGGCCCAAGTTCACTTAAAAAGGAGATCAACAATGAAA
                     GCAATTTTCGTACTGAAACATCTTAATCATGCGAGCTTTTCACTGGCGTTGTTCCCAT
                     CCTGGTCGAGCTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGC
                     GAGGGCGATGCCACCTACGGCAAGCTGACCCTGAAGTTCATCTGCACCACCGGCAAGC
                     TGCCCGTGCCCTGGCCCACCCTCGTGACCACCCTGACCTACGGCGTGCAGTGCTTCAG
                     CCGCTACCCCGACCACATGAAGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGC
                     TACGTCCAGGAGCGCACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCG
                     AGGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGCATCGACTT
                     CAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGAGTACAACTACAACAGCCACAAC
                     GTCTATATCATGGCCGACAAGCAGAAGAACGGCATCAAGGTGAACTTCAAGATCCGCC
                     ACAACATCGAGGACGGCAGCGTGCAGCTCGCCGACCACTACCAGCAGAACACCCCCAT
                     CGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCACCCAGTCCGCCCTG
                     AGCAAAGACCCCAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTCGTGACCGCCG
                     CCGGGAT"
                     /label="TAAGGGCCCAAGTTCACTTAAAAAGGAGATCAACAATGAAAGCAATTTTC
                     GTACTGAAACATCTTAATCATGCGAGCTTTTCACTGGCGTTGTTCCCATCCTGGTCGA
                     GCTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGAT
                     GCCACCTACGGCAAGCTGACCCTGAAGTTCATCTGCACCACCGGCAAGCTGCCCGTGC
                     CCTGGCCCACCCTCGTGACCACCCTGACCTACGGCGTGCAGTGCTTCAGCCGCTACCC
                     CGACCACATGAAGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAG
                     GAGCGCACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGT
                     TCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGA
                     CGGCAACATCCTGGGGCACAAGCTGGAGTACAACTACAACAGCCACAACGTCTATATC
                     ATGGCCGACAAGCAGAAGAACGGCATCAAGGTGAACTTCAAGATCCGCCACAACATCG
                     AGGACGGCAGCGTGCAGCTCGCCGACCACTACCAGCAGAACACCCCCATCGGCGACGG
                     CCCCGTGCTGCTGCCCGACAACCACTACCTGAGCACCCAGTCCGCCCTGAGCAAAGAC
                     CCCAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTCGTGACCGCCGCCGGGAT"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    558..>584
                     /Source="referenceToDocument: urn:local:.:3y-824h256"
                     /label="deGFP (21) (concatenated sequence 3)"
                     /note="Geneious type: Concatenated sequence"
     CDS             585..>1298
                     /created_by="User"
                     /label="sfGFP BBa Safe"
     primer_bind     926..942
                     /Sequence="TGAAGGCGATACCCTGG"
                     /Hairpin_Tm="40.6"
                     /Tm="55.4"
                     /Self_Dimer_Tm="None"
                     /%GC="58.8"
                     /modified_by="User"
                     /label="Oligo_82f"
     misc_feature    complement(1086..1100)
                     /Sequence="CTCCACGTTGTGGCG"
                     /Hairpin_Tm="51.6"
                     /Tm="54.6"
                     /Self_Dimer_Tm="11.1"
                     /%GC="66.7"
                     /modified_by="User"
                     /label="Oligo_82r"
                     /note="Geneious type: primer_bind_reverse"
     misc_difference 1299..1316
                     /created_by="User"
                     /label="His Tag"
     misc_feature    1300..1316
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     misc_feature    <1317..34
                     /label="Digestion of piece 1 ext by BsaI - Fragment 2
                     (concatenated sequence 1)"
                     /note="Geneious type: Concatenated sequence"
     misc_feature    1318
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     terminator      1327..1356
                     /note="T500"
                     /label="T500 terminator"
     conflict        1393..1417
                     /created_by="User"
                     /label="area for IA_v1, v2"
     conflict        1396..1410
                     /created_by="User"
                     /label="area for IB_v1, v2"
     modified_base   1445
                     /created_by="User"
                     /label="c->g"
     misc_feature    1445
                     /Original_Bases="C"
                     /label="C"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    1484
                     /Original_Bases="G"
                     /label="G"
                     /note="Geneious type: Editing History Replacement"
     misc_difference <1485..>1506
                     /created_by="User"
                     /label
     misc_feature    1507
                     /Original_Bases="G"
                     /label="G"
                     /note="Geneious type: Editing History Replacement"
     rep_origin      complement(1650..2332)
                     /ApEinfo_fwdcolor="gray50"
                     /ApEinfo_revcolor="gray50"
                     /label="ColE1 origin"
     misc_feature    1862^1863
                     /Original_Bases="A"
                     /label="A"
                     /note="Geneious type: Editing History Deletion"
     misc_feature    1864
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     misc_feature    2109
                     /Original_Bases="G"
                     /label="G"
                     /note="Geneious type: Editing History Replacement"
     CDS             complement(2427..3287)
                     /ApEinfo_fwdcolor="yellow"
                     /ApEinfo_revcolor="yellow"
                     /modified_by="User"
                     /label="AmpR"
     modified_base   2568
                     /created_by="User"
                     /label="a->t"
     misc_feature    2568
                     /Original_Bases="A"
                     /label="A"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    2818
                     /created_by="User"
                     /label="A->G unique mut. not inherited from 21p"
     misc_feature    2818
                     /Original_Bases="A"
                     /label="A"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    3044
                     /created_by="User"
                     /label="nonsilenced mutation Ile -> Val"
     misc_feature    3044
                     /Original_Bases="T"
                     /label="T"
                     /note="Geneious type: Editing History Replacement"
     modified_base   3487
                     /created_by="User"
                     /label="c->g"
     misc_feature    3487
                     /Original_Bases="C"
                     /label="C"
                     /note="Geneious type: Editing History Replacement"
     modified_base   3589
                     /created_by="User"
                     /label="a->t"
     misc_feature    3589
                     /Original_Bases="A"
                     /label="A"
                     /note="Geneious type: Editing History Replacement"
     modified_base   3627
                     /created_by="User"
                     /modified_by="User"
                     /label="a->t"
     misc_feature    3627
                     /Original_Bases="A"
                     /label="A"
                     /note="Geneious type: Editing History Replacement"
     conflict        3693..3709
                     /created_by="User"
                     /label="area for IA_vb1, vb2"
     conflict        3695..3712
                     /created_by="User"
                     /label="area for IA_vA1, IA_vA2"
     misc_feature    3749..3803
                     /ApEinfo_fwdcolor="#ff0000"
                     /ApEinfo_revcolor="green"
                     /label="OR2-OR1-Pr"
     promoter        3768..3773
                     /created_by="User"
                     /label="-35"
     promoter        3791..3796
                     /created_by="User"
                     /modified_by="User"
                     /label="-10"
ORIGIN      
        1 aataattttg tttaacttta agaaggagat atacaatgga cgaagttgat ggtccgccaa
       61 tgttgtccgg tctgcttgct cgtctggtta agttgcttct gggccgtcac ggttctgcgc
      121 tgcactggcg tgctgcaggc gctgccaccg tccttctggt gatcgtactg cttgctggtt
      181 cctacttggc ggttctggcg gaagcggatg atccgaccag cggctttagc agcattccgg
      241 acgccttgtg gtggtctgtc gataccgcta ctaccgtggg ttacggcgac ttgtatccgg
      301 tgaccctttg gggtcgtctg gtagcggttg tcgttatggt ggctggcatc acctccttcg
      361 gtctggtaac tgctgccctt gctacttggt ttgtaggccg cgaacaagag cgtcgtggtc
      421 atttcgttcg tcactctgaa aaagcggctg aggaggcgta cacccgtact acccgtgctc
      481 tgcacgagcg ctttgaccgt ctggaacgta tgcttgacga taaccgtcgt ggtgaaaacc
      541 tgtacttcca gtctggtaac ctgtacttcc agtctggtgg tgctatgcgt aaaggagaag
      601 aacttttcac tggtgtcgtc cctattctgg tggaactgga tggtgatgtc aacggtcata
      661 agttttccgt gcgtggcgag ggtgaaggtg acgcaactaa tggtaaactg acgctgaagt
      721 tcatctgtac tactggtaaa ctgccggtac cttggccgac tctggtaacg acgctgactt
      781 atggtgttca gtgctttgct cgttatccgg accatatgaa gcagcatgac ttcttcaagt
      841 ccgccatgcc ggaaggctat gtgcaggaac gcacgatttc ctttaaggat gacggcacgt
      901 acaaaacgcg tgcggaagtg aaatttgaag gcgataccct ggtaaaccgc attgagctga
      961 aaggcattga ctttaaagaa gatggcaata tcctgggcca taagctggaa tacaatttta
     1021 acagccacaa tgtttacatc accgccgata aacaaaaaaa tggcattaaa gcgaatttta
     1081 aaattcgcca caacgtggag gatggcagcg tgcagctggc tgatcactac cagcaaaaca
     1141 ctccaatcgg tgatggtcct gttctgctgc cagacaatca ctatctgagc acgcaaagcg
     1201 ttctgtctaa agatccgaac gagaaacgcg atcatatggt tctgctggag ttcgtaaccg
     1261 cagcgggcat cacgcatggt atggatgaac tgtacaaaca tcatcatcat catcattgaa
     1321 ctcgagcaaa gcccgccgaa aggcgggctt ttctgtgtcg accgatgccc ttgagagcct
     1381 tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga
     1441 ctgtgttctt tatcatgcaa ctcgtaggac aggtgccggc agctctcttc cgcttcctcg
     1501 ctcacttact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     1561 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     1621 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     1681 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1741 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1801 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     1861 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     1921 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     1981 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2041 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2101 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2161 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2221 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2281 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2341 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2401 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2461 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     2521 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgtga cccacgctca
     2581 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     2641 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     2701 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     2761 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttgca
     2821 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     2881 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     2941 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3001 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg
     3061 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3121 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3181 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3241 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3301 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3361 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     3421 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     3481 tttcgtgttc aagaattctg gcgaatcctc tgaccagcca gaaaacgacc tttctgtggt
     3541 gaaaccggat gctgcaattc agagcgccag caagtggggg acagcagatg acctgaccgc
     3601 cgcagagtgg atgtttgaca tggtgatgac tatcgcacca tcagccagaa aaccgaattt
     3661 tgctgggtgg gctaacgata tccgcctgat gcgtgaacgt gacggacgta accaccgcga
     3721 catgtgtgtg ctgttccgct gggcatgctg agctaacacc gtgcgtgttg acaattttac
     3781 ctctggcggt gataatggtt gcagctagc
//
