LOCUS       pCAD183_gCAD_VLTILLK_pET        6698 bp    DNA     circular UNA 22-JUL-2020
DEFINITION  Golden Gate/Type IIS Cloning ligation product.
ACCESSION   urn.local...d3-c3nn1w9
VERSION     urn.local...d3-c3nn1w9
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     primer_bind     267..281
                     /Sequence="ATGCGTCCGGCGTAG"
                     /%GC="66.7"
                     /Transferred_From="pET-up"
                     /Transferred_Similarity="100.00%"
                     /label="pET-up"
     promoter        309..327
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /label="T7 promoter"
     primer_bind     312..336
                     /created_by="primer3"
                     /%GC="44.0"
                     /Tm="59.7"
                     /Hairpin_Tm="35.0"
                     /Self_Dimer_Tm="2.2"
                     /Sequence="TACGACTCACTATAGGGGAATTGTG"
                     /Transferred_From="pOPINE_fwd"
                     /Transferred_Similarity="100.00%"
                     /label="pOPINE_fwd"
     misc_feature    326
                     /created_by="kbu07"
                     /label="Transcription start"
                     /note="Geneious type: Transcription start"
     protein_bind    328..352
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
                     /label="lac operator"
     RBS             367..389
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
                     /label="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     MoClo           395..398
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
                     /label="A_ATG"
     CDS             <399..1841
                     /Genetic_code="Standard"
                     /Reading_frame=1
                     /created_by="ORF Finder"
                     /modified_by="kbu07"
                     /label="Strep-TEV-gCAD"
     primer_bind     399..417
                     /Sequence="GCTAGCTGGAGCCACCCGC"
                     /%GC="73.7"
                     /Extension="GCGGGTACCGGTCTCAAATG"
                     /Transferred_From="cCAD-For1-GoldMut"
                     /Transferred_Similarity="100.00%"
                     /label="cCAD-For1-GoldMut"
     CDS             <401..>464
                     /codon_start=2
                     /label="StrepTag-TEV-NotI"
     protein_binding 405..428
                     /ApEinfo_fwdcolor="#00ff00"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}"
                     /ApEinfo_revcolor="#00ff00"
                     /label="StreptagII"
     protein_binding 435..455
                     /ApEinfo_fwdcolor="#ffff00"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}"
                     /ApEinfo_revcolor="#ffff00"
                     /label="TEV-Site"
     misc_feature    463..464
                     /created_by="User"
                     /label="GC"
                     /note="Geneious type: Gibson Primer Extension"
     misc_feature    464..465
                     /Original_Bases="AUGAUGCUC"
                     /label="AUGAUGCUC"
                     /note="Geneious type: Editing History Deletion"
     misc_difference 917
                     /note="G changed to A in pCAD29_hIRG1_4-461_pvp008"
                     /label="difference"
     ligation        1601..1604
                     /label="Ligation"
     primer_bind     complement(1795..1820)
                     /%GC="38.5"
                     /Hairpin_Tm="28.8"
                     /Self_Dimer_Tm="8.6"
                     /Tm="59.7"
                     /Pair_Dimer_Tm="None"
                     /Sequence="AGTTAACACAGAACAGTCTTCTAGGT"
                     /created_by="primer3"
                     /Product_Size=1422
                     /Extension="CACCACAGGTCTCGACCTTCAGGGTCCTTTGAGAAGTAT"
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="gCAD_CAD177_VLTILLK_R"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:e9-c3nq247"">gCAD_CAD177_VLTILLK_R</a>"
                     /label="gCAD_CAD177_VLTILLK_R"
     misc_feature    1841^1842
                     /Original_Bases="AGGT"
                     /label="AGGT"
                     /note="Geneious type: Editing History Deletion"
     MoClo           1842..1845
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
                     /label="A_GGT"
     ligation        1842..1845
                     /label="Ligation"
     primer_bind     complement(1933..1956)
                     /created_by="primer3"
                     /%GC="54.2"
                     /Tm="65.5"
                     /Hairpin_Tm="40.4"
                     /Self_Dimer_Tm="17.6"
                     /Sequence="TGCTAGTTATTGCTCAGCGGTGGC"
                     /Transferred_From="RSF_rev"
                     /Transferred_Similarity="100.00%"
                     /label="RSF_rev"
     terminator      1951..1998
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
                     /label="T7 terminator"
     rep_origin      2035..2490
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
                     /label="f1 ori"
     CDS             complement(2583..3398)
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
                     /label="KanR"
     rep_origin      3520..4108
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /label="ori"
     misc_feature    4294..4436
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /label="bom"
     CDS             complement(4538..4729)
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
                     /label="rop"
     CDS             complement(5538..6620)
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
                     /label="lacI"
     promoter        complement(6621..6698)
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
                     /label="lacI promoter"
ORIGIN      
        1 cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
       61 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca
      121 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
      181 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
      241 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc
      301 cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga
      361 aataattttg tttaacttta agaaggagat atacaatggc tagctggagc cacccgcagt
      421 tcgaaaaagt cgacgagaac ctctacttcc agggaggcgg ccgcaagtct atcacagaaa
      481 gctttgccac agcaatccat ggcttgaaag tgggacacct gacagatcgt gttattcaga
      541 ggagcaagag gatgattcta gacactctgg gtgctgggtt cctgggaacc actacggaag
      601 tgtttcacat agccagccaa tatagcaaga tctacagttc caacatatcc agcactgttt
      661 ggggtcagcc agacatcagg ctcccgccca catatgctgc ttttgtgaac ggtgtggcta
      721 ttcactccat ggattttgat gacacgtggc accctgccac ccacccttct ggggctgtcc
      781 ttcctgtcct cacagcttta gcagaagccc tgccaaggag tccaaagttt tctggccttg
      841 acctgctgct ggctttcaat gttggtattg aagtgcaagg ccgattactg catttcgcca
      901 aggaggccaa tgacatacca aagagattcc atcccccttc cgtggtagga acgttgggta
      961 gtgctgctgc tgcatccaag tttttaggac ttagctcgac aaagtgccga gaagctctgg
     1021 ccattgctgt ttcccatgct ggggcaccca tggccaatgc tgccacccag accaagcccc
     1081 tccacattgg caatgctgcc aagcatggga tagaagctgc atttttggca atgttgggtc
     1141 tgcaaggaaa caagcaggtc ttggacttgg aggcaggatt tggggccttt tatgccaact
     1201 attccccaaa agtccttcca agcctggctt cctacagttg gctgctggac cagcaggacg
     1261 tggcctttaa gcgttttcct gcacatttat ctacccactg ggtggcagac gcagctgcat
     1321 ctgtgagaaa gcaccttgta gcagagagag ccctgcttcc aactgactac attaagagaa
     1381 ttgtgctcag gataccaaat gtccagtatg taaacaggcc ctttccagtt tcggagcatg
     1441 aagcccgtca ttcattccag tatgtggcct gtgccatgct gcttgatggt ggcatcactg
     1501 tcccctcatt ccatgaatgc cagatcaaca ggccacaggt gagagagctg ctcagtaagg
     1561 tggagctgga gtaccctccg gacaacttgc caagcttcaa caccctgtac tgtgaaataa
     1621 gtgtcaccct caaggatgga gccaccttca cagatcgctc tgataccttc tatgggcact
     1681 ggagaaaacc actgagccag gaggacctag aggaaaagtt cagagccaat gcctccaaga
     1741 tgctgtcctg ggacacagtg gaaagcctta taaagatagt cgaaaatcta gaagacctag
     1801 aagactgttc tgtgttaact atacttctca aaggaccctg aaggttagta agcggccgca
     1861 ctcgagcacc accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct
     1921 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     1981 gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa tgggacgcgc
     2041 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2101 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2161 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2221 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     2281 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     2341 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     2401 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     2461 attttaacaa actagtaacg tttacaattt caggtggcac ttttcgggga aatgtgcgcg
     2521 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat
     2581 tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     2641 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     2701 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     2761 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     2821 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     2881 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     2941 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     3001 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tgttttcacc tgaatcagga
     3061 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
     3121 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     3181 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     3241 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     3301 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     3361 ggcctagagc aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
     3421 atgtaagcag acagttttat tgttcatgac caaaatccct taacgtgagt tttcgttcca
     3481 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3541 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3601 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3661 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3721 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3781 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3841 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3901 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3961 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4021 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4081 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4141 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     4201 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     4261 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     4321 tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc
     4381 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     4441 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     4501 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     4561 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     4621 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     4681 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     4741 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
     4801 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
     4861 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
     4921 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
     4981 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
     5041 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
     5101 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
     5161 tcctcaacga caggagcacg atcatgcgca cccgtggggc cgccatgccg gcgataatgg
     5221 cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt
     5281 gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt
     5341 cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga
     5401 agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg
     5461 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
     5521 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5581 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
     5641 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     5701 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     5761 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata
     5821 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
     5881 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
     5941 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
     6001 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
     6061 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
     6121 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
     6181 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
     6241 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
     6301 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
     6361 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
     6421 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
     6481 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     6541 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     6601 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     6661 gccataccgc gaaaggtttt gcgccattcg atggtgtc
//
