LOCUS       pBK_Ma_Pyl_RS_WT        2854 bp DNA     circular SYN 20-MAY-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 2854)
  AUTHORS   Savanna's PC
  TITLE     Direct Submission
  JOURNAL   Exported Jan 26, 2023 from SnapGene 6.2.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..2854
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     misc_feature    2..14
                     /label=New Feature(1)
                     /note="ApEinfo_fwdcolor=#ff8000
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_label=New Feature
                     ApEinfo_revcolor=green"
                     /note="color: #ff8000"
     primer_bind     21..47
                     /label=pBK 50 bp upstream
                     /note="color: black; sequence: cgcttataagatcatacgccgttatac;
                     added: 2020-03-11"
     promoter        21..32
                     /label=GlnS p
                     /note="color: #ff58aa"
     promoter        30..36
                     /label=constitutive glutamine promotor (from E.coli gl
                     /label=constitutive\glutamine\promotor\(from\E.coli\glnS\g
                     /note="ApEinfo_fwdcolor=pink
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_revcolor=pink
                     vntifkey=30"
                     /note="color: #ffc0cb"
     primer_bind     34..55
                     /label=MaHCO_40bb_from_start_for
                     /note="color: black; sequence: atacgccgttatacgttgttta; 
                     added: 2019-08-09"
     RBS             63..68
                     /label=rbs
                     /note="ApEinfo_fwdcolor=pink
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_revcolor=pink
                     vntifkey=32"
                     /note="color: #ffc0cb"
     misc_feature    71..72
                     /label=NdeI
                     /note="color: #dff208"
     CDS             74..907
                     /codon_start=1
                     /label=HCO Ma bocK PylRS
                     /note="color: #993366"
                     /translation="MTVKYTDAQIQRLREYGNGTYEQKVFEDLASRDAAFSKEMSVAST
                     DNEKKIKGMIANPSRHGLTQLMNDIADALVAEGFIEVRTPIFISKDALARMTITEDKPL
                     FKQVFWIDEKRALRPMLAPNLYSVMRDLRDHTDGPVKIFEMGSCFRKESHSGMHLEEFT
                     MLNLVDMGPRGDATEVLKNYISVVMKAAGLPDYDLVQEESDVYKETIDVEINGQEVCSA
                     AVGPHYLDAAHDVHEPWSGAGFGLERLLTIREKYSTVKKGGASISYLNGAKINSG"
     primer_bind     complement(76..98)
                     /label=Ma lib vector flank 5' (tgcc)
                     /note="color: black; sequence: 
                     actgcggtctcGGGCATCTGTGTATTTCACTGTC; added: 2020-09-11"
     primer_bind     77..100
                     /label=Ma to pET28-bdSUMO-for
                     /note="color: black; sequence: 
                     GCGATGCTGCATCAGACCGGCGGATCCGGTACAGTGAAATACACAGATGCCCAG; 
                     added: 2021-11-16"
     primer_bind     93..126
                     /label=Ma lib flank for
                     /note="color: black; sequence: 
                     acgttggtctcATGCCCAGATCCAGCGCCTGCGGGAGTATGGCAA; added: 
                     2020-03-11"
     regulatory      249..258
                     /label=Ma RS
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     primer_bind     402..442
                     /label=5'P_overlap_ext_for
                     /note="color: black; sequence: 
                     GGATCGATGAGAAGAGGGCTCTGAGGCCCATGCTCGCCCCC; added: 
                     2020-09-21"
     primer_bind     complement(417..436)
                     /label=MaHCO_L121_rev
                     /note="color: black; sequence: GAGCATGGGCCTCAGAGCCC; added:
                     2019-08-20"
     primer_bind     420..479
                     /label=Ma D3 lib 125-126-129 for(1)
                     /note="color: black; sequence: 
                     CTCTGAGGCCCATGCTCGCCCCCAACnnnnnnTCCGTGnnnCGGGACCTGcgcGACCAC
                     A; added: 2020-03-11"
     primer_bind     421..476
                     /label=Ma S2 125 ndt for
                     /note="color: black; sequence: 
                     TCTGAGGCCCATGCTCGCCCCCAACndtTACTCCGTGATGCGGGACCTGcgcGACC; 
                     added: 2020-09-11"
     primer_bind     complement(421..445)
                     /label=PCR 1 rev
                     /note="color: black; sequence: GTTGGGGGCGAGCATGGGCCTCAGA; 
                     added: 2020-03-11"
     primer_bind     complement(424..482)
                     /label=Ma Y126A-M129L rev
                     /note="color: black; sequence: 
                     CCGTGTGGTCgcgCAGGTCCCGCAGCACGGAGGCCAGGTTGGGGGCGAGCATGGGCCTC
                     ; added: 2020-05-12"
     primer_bind     424..482
                     /label=Ma Y126A-M129L for
                     /note="color: black; sequence: 
                     GAGGCCCATGCTCGCCCCCAACCTGGCCTCCGTGCTGCGGGACCTGcgcGACCACACGG
                     ; added: 2020-05-12"
     primer_bind     complement(448..502)
                     /label=Bsa(1) remove rev
                     /note="color: black; sequence: 
                     GAAAATTTTCACAGGGCCGTCCGTGTGGTCgcgCAGGTCCCGCATCACGGAGTAC; 
                     added: 2020-03-11"
     primer_bind     448..502
                     /label=Bsa(1) remove for
                     /note="color: black; sequence: 
                     GTACTCCGTGATGCGGGACCTGcgcGACCACACGGACGGCCCTGTGAAAATTTTC; 
                     added: 2020-02-27"
     primer_bind     539..598
                     /label=Ma D3-S2 lib 166-168 for(1)
                     /note="color: black; sequence: 
                     GGAATGCACCTGGAGGAGTTCACCATGCTGnnnCTGnnnGACATGGGGCCAAGAGGAGA
                     T; added: 2020-03-11"
     primer_bind     complement(540..568)
                     /label=PCR 2 rev
                     /note="color: black; sequence: 
                     CAGCATGGTGAACTCCTCCAGGTGCATTC; added: 2020-03-11"
     primer_bind     complement(544..596)
                     /label=MaRS_N166TAA_rev
                     /note="color: black; sequence: 
                     CTCCTCTTGGCCCCATGTCCACCAGTTACAGCATGGTGAACTCCTCCAGGTGC; 
                     added: 2020-12-24"
     primer_bind     544..596
                     /label=MaRS_N166TAA_for
                     /note="color: black; sequence: 
                     GCACCTGGAGGAGTTCACCATGCTGTAACTGGTGGACATGGGGCCAAGAGGAG; 
                     added: 2020-12-24"
     primer_bind     673..721
                     /label=Ma Bsa(2) for
                     /note="color: black; sequence: 
                     GGAAGAGAGCGATGTCTACAAAGAaACCATTGATGTGGAGATCAATGGC; added: 
                     2020-02-27"
     primer_bind     complement(673..721)
                     /label=Bsa(2) remove rev
                     /note="color: black; sequence: 
                     GCCATTGATCTCCACATCAATGGTtTCTTTGTAGACATCGCTCTCTTCC; added: 
                     2020-03-11"
     primer_bind     complement(719..778)
                     /label=Ma S2 223 ndt rev
                     /note="color: black; sequence: 
                     CACGTCGTGGGCGGCGTCCAGGTAGTGGGGGCCCACahnAGCAGAGCACACCTCCTGGC
                     C; added: 2020-09-11"
     primer_bind     755..814
                     /label=Ma S2 239 ndt for
                     /note="color: black; sequence: 
                     TACCTGGACGCCGCCCACGACGTGCATGAACCCndtAGTGGAGCGGGCTTTGGCCTGGA
                     G; added: 2020-09-11"
     primer_bind     complement(801..840)
                     /label=5'P_overlap_ext_rev
                     /note="color: black; sequence: 
                     TACTTTTCTCTTATGGTCAGCAGCCTCTCCAGGCCAAAGC; added: 
                     2020-09-21"
     primer_bind     complement(872..898)
                     /label=Ma to pET28 rev
                     /note="color: black; sequence: 
                     GATCTCAGTGGTGGTGGTGGTGGTGCTCGAGttaGTTGATCTTGGCTCCATTCAAGTAG
                     GA; added: 2021-11-16"
     primer_bind     complement(892..938)
                     /label=Ma lib flank rev
                     /note="color: black; sequence: 
                     actgtGGTCTCaggcaatttagcgtttgaaactgcagGCTTCAGCCGCTGTTGATC; 
                     added: 2020-03-11"
     misc_feature    912..916
                     /label=PstI
                     /note="color: #dff208"
     primer_bind     complement(940..966)
                     /label=pBK 50 bp from end
                     /note="color: black; sequence: atcatgtaggcctgataagcgtagcgc;
                     added: 2020-03-11"
     primer_bind     952..981
                     /label=Ma lib vector flank 3' (cagg)
                     /note="color: black; sequence: 
                     agctcGGTCTCtcaggcctacatgatctctgcaatatattg; added: 
                     2020-09-11"
     misc_feature    954..959
                     /label=StuI
                     /note="color: #dff208"
     primer_bind     complement(997..1018)
                     /label=Rev seq primer
                     /note="color: black; sequence: gtgaacgccttatccggcctac"
     terminator      1050..1074
                     /label=terminator (from E.coli glnS glutamine syntheta
                     /label=terminator\(from\E.coli\glnS\glutamine\synthetase)
                     /note="ApEinfo_fwdcolor=pink
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_revcolor=pink
                     vntifkey=43"
                     /note="color: #ffc0cb"
     rep_origin      1147..1747
                     /label=ColE1 Origin
                     /label=ColE1\Origin
                     /note="ApEinfo_fwdcolor=pink
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_revcolor=pink
                     vntifkey=33"
                     /note="color: #ffc0cb"
     misc_feature    1937..2752
                     /label=KAN
                     /label=misc_marker
                     /note="ApEinfo_fwdcolor=pink
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_revcolor=pink
                     vntifkey=22"
                     /note="color: #ffc0cb"
     primer_bind     join(2839..2854,1..6)
                     /label=Forward seq primer
                     /note="color: black; sequence: gctgagttgaaggatcctcggg"
ORIGIN
        1 ctcgggttgt cagcctgtcc cgcttataag atcatacgcc gttatacgtt gtttacgctt
       61 tgaggaatcc catatgacag tgaaatacac agatgcccag atccagcgcc tgcgggagta
      121 tggcaacggc acctatgagc agaaggtgtt tgaagatctg gcctctagag atgcagcctt
      181 ctccaaggag atgtccgtgg cttccacaga caacgagaaa aagatcaagg gcatgattgc
      241 caaccccagc cgccatgggc tgacccagct gatgaatgac atcgccgacg ccctggtggc
      301 cgagggcttc atcgaggtca ggacccccat cttcatttct aaggacgcgc tggctcgcat
      361 gaccatcacc gaggacaagc ccctgttcaa gcaggtgttc tggatcgatg agaagagggc
      421 tctgaggccc atgctcgccc ccaacctgta ctccgtgatg cgggacctgc gcgaccacac
      481 ggacggccct gtgaaaattt tcgaaatggg ctcctgcttt aggaaagaaa gccacagcgg
      541 aatgcacctg gaggagttca ccatgctgaa tctggtggac atggggccaa gaggagatgc
      601 cacagaagtg ctgaagaact acatctcagt ggtcatgaag gctgctggac tgcccgacta
      661 tgatttggtg caggaagaga gcgatgtcta caaagaaacc attgatgtgg agatcaatgg
      721 ccaggaggtg tgctctgctg ccgtgggccc ccactacctg gacgccgccc acgacgtgca
      781 tgaaccctgg agtggagcgg gctttggcct ggagaggctg ctgaccataa gagaaaagta
      841 cagcactgtg aagaaaggcg gcgcctccat ctcctacttg aatggagcca agatcaacag
      901 cggctgaagc ctgcagtttc aaacgctaaa ttgcctgatg cgctacgctt atcaggccta
      961 catgatctct gcaatatatt gagtttgcgt gcttttgtag gccggataag gcgttcacgc
     1021 cgcatccggc aagaaacagc aaacaatcca aaacgccgcg ttcagcggcg ttttttctgc
     1081 ttttcttcgc gaattaattc cgcttcgcac atgtgagcaa aaggccagca aaaggccagg
     1141 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1201 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1261 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1321 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     1381 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1441 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     1501 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     1561 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     1621 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     1681 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     1741 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     1801 aacgaaaact cacgttaagg gattttggtc atgagttgtg tctcaaaatc tctgatgtta
     1861 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     1921 taatacaagg ggtgttatga gccatattca acgggaaacg tcttgctcga ggccgcgatt
     1981 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     2041 atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa
     2101 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     2161 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     2221 gttactcacc actgcgatcc ccgggaaaac agcattccag gtattagaag aatatcctga
     2281 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     2341 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     2401 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     2461 tgaacaagtc tggaaagaaa tgcataagct tttgccattc tcaccggatt cagtcgtcac
     2521 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     2581 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     2641 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     2701 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     2761 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcggctttgt
     2821 tgaataaatc gaacttttgc tgagttgaag gatc
//
