LOCUS       pSGAb/Evol-Tac-AbWRS(AG 6417 bp ds-DNA     circular     23-OCT-2024
DEFINITION  .
KEYWORDS    "accession:addgene_121999_239475"
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
COMMENT     GENERAL Depositing Lab: Quanjiang Ji Publication: Wang et al Cell
            Chem Biol. 2019 Sep 17. pii: S2451-9456(19)30277-6. doi:
            10.1016/j.chembiol.2019.09.003. Purpose: A sgRNA expression plasmid
            for genome editing in Acinetobacter baumannii BACKBONE Backbone
            size w/o insert (bp): 6073 Modifications to backbone: pSGAb-km was
            constructed by combining two different replication regions:
            pMB1_origin (from pUC57 vector) and WH1266_origin (from pWH1266
            vector). Vector backbone: pUC57 and pWH1266 Vector type: CRISPR
            CLONING INFORMATION 5′ sequencing primer: Unknown Cloning method:
            Gibson Cloning GROWTH IN BACTERIA Bacterial Resistance(s):
            Kanamycin Copy number: High Copy Growth Strain(s): DH5alpha Growth
            Temperature: 37°C Growth instructions: Selectable Marker: 50 μg/mL
            kanamycin in both E.coli DH5alpha and Acinetobacter baumannii
            GENE/INSERT Gene/Insert name: none gRNA/shRNA sequence: unknown
FEATURES             Location/Qualifiers
     primer_bind     complement(29..51)
                     /label="pGEX 3'"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    187..1519
                     /label="WH1266 Ori"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     CDS             complement(1583..1651)
                     /label="LacZ alpha"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    complement(1672..1703)
                     /label="PIPE primer?"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          complement(1672..1718)
                     /label="pEvol-pSGAb-PIPE2-R"
                     /note="sequence: cccggtcatcaatcaccctggcgttacccaacttaatcgccttgcag"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     primer          1690..1732
                     /label="pEvol-pSGAb-PIPE2-F"
                     /note="sequence: gggtaacgccagggtgattgatgaccgggagctcacaaacaag"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer          1704..1732
                     /label="pEvol-Tac-rrnB-R"
                     /note="sequence: tgattgatgaccgggagctcacaaacaag"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    complement(1730..1887)
                     /label="rrnB terminator"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer          complement(1894..1920)
                     /label="EcW-sqiF"
                     /note="sequence: CAGAACGCAGAAGCGGTCTGATAAAAC"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     misc_feature    complement(1998..3011)
                     /label="A. baumannii tryptophanyl tRNA synthetase"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     CDS             complement(2001..3011)
                     /label="Translation 3011-2001"
                     /translation="MSNVDQRPIILAGDRPTGQLHLGHFVGSLRSRVGLQDSHHQHLLLADAQALTDNADNPDKVRRNILEVALDYLAVGIDPTKTTICVQSCLPALNELTMLYLNFVTVARLERNPTIKSEIQMRGFERDIPAGFLCYPVAQAADITAFKATVGPCGEDQIPMIEQTNEIVRRVNRQIGQDLLPECKALLSNMARLPGFDGKAKMSKSLGNTIVLNASDKDIKKAVNAMYTDPNHLRIEDPGQVEGNIVFTYLDAFDPNKEEVEELKAHYRRGGLGDGTVKKRLEGVLKELITPIRERREELAKDPDYIMDVLRQGTDKCRIITQQTLDEVKDGLGLFKF"
     primer          complement(2521..2579)
                     /label="BaumanniiTrp-151G,152C-F"
                     /note="sequence: GCTTTTAAAGCGACTGTAGGACCATGCGGCGAAGATCAAATTCCAATGATTGAGCAAAC"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     primer          2551..2607
                     /label="BaumanniiTrp-151G,153C-R"
                     /note="sequence: CCGCATGGTCCTACAGTCGCTTTAAAAGCCGTAATATCAGCTGCTTGTGCTACAGGG"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     misc_feature    complement(2553..2555)
                     /label="V153>C"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     misc_feature    complement(2559..2561)
                     /label="V151>G"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     misc_feature    complement(2976..2978)
                     /label="12T>A"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    complement(3022..3097)
                     /label="TacI promoter"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     misc_feature    complement(3049..3049)
                     /label=3
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     misc_feature    complement(3059..3065)
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    complement(3059..3087)
                     /label="tacI promoter"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     misc_feature    complement(3082..3087)
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     misc_feature    complement(3213..4292)
                     /label="lacI"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    complement(3384..3384)
                     /label="mutated to T in 14-8"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     misc_feature    complement(3385..3385)
                     /label="mutated to T in 9-2"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     misc_feature    complement(3424..3424)
                     /label="mutated to G in 9-1"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     primer          complement(4474..4531)
                     /label="pEvol-pSGAb-PIPE-R"
                     /note="sequence: gagtgagctgataccgctcgccgcctcatgtttgacagcttatcatcgataagcttgc"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer          4496..4536
                     /label="pEvol-pSGAb-PIPE-F"
                     /note="sequence: gtcaaacatgaggcggcgagcggtatcagctcactcaaagg"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     misc_feature    4508..4536
                     /label="PIPE primer F?"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     rep_origin      complement(4625..5307)
                     /label="ColE1 origin"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     rep_origin      complement(4643..5231)
                     /label="ori"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer_bind     complement(4723..4742)
                     /label="pBR322ori-F"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     CDS             complement(5402..6217)
                     /label="KanR"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer_bind     6125..6144
                     /label="Kan-R"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     promoter        complement(6218..6322)
                     /label="AmpR promoter"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer_bind     6390..6408
                     /label="pBRforEco"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataggat tttaacattt tgcgttgttc caaaagttat caacagccta gaacgtcata
      241 ggaagcgatt acagacactt tatagctatc agcatgggaa cataagggca ggatgaaata
      301 tgggtcttaa aacgcaaatg gtgaggtttt agaggtattt tttgaagatg attaaggcgg
      361 tttgttttta aaatttttgg cggctctcag gctgcttaca ttttaaccag ttcagtgaaa
      421 agttcttttt cagcaaattt ctgtttagca ccatagctaa aacttgcgtg gaacatatta
      481 agaattgacc gaaatgacac aatctcaatt atattttttt tgaaaagttt tctttatcaa
      541 atattttaaa tcattgattt atatataagt atgcattcat tttaataatt aatctttatt
      601 taacaatgat ttatctatat tcaattgttt aattattctt actaatatta tctctatatc
      661 aatatttttt atttaaaaac atatgtttag tagtgctttt gattaaagta ccagagggag
      721 ggagcagagc tgaatgggaa atactcaccc tagagcgatt cttaaaaatc accctaaagt
      781 attcccattc gatgtaccgt cggtcggtcg ctttcgcatc agggatgaca tcactgtatc
      841 aagctgccac tgttatgatt acgattgata gcaccgcctg aacacgctca taaccgccaa
      901 ttaaatgact actcattgcg tccgctactc tgttcagttc ccctagtaat agcgtttttc
      961 cgatgtgtgc tagcgtcact gtacctcatc acccacacat ggacagattg agttacagac
     1021 attggctaaa tttttggggt ctatgcttga caaagcgagt taaaacctta gaatttaaga
     1081 caggtacatt aagcccctgt ggtgaaatca ttagcggtcg tcaaacctta atgctttcga
     1141 ttgccgatat gtcagtatcg aagatctgta ccccataaat tacggtaaag ccccaagcaa
     1201 ttgcaagggg cttttatctt ttttaacaaa aaaaatttat aaatcaggat tttataacac
     1261 taaataatcc aaagtacatg agtaagtcat gaccactcct gcgatgtgtg tagcactctg
     1321 agtatccgta ttatatcagt gatgtcatat acaaccacat aatgcggatg tatcactaac
     1381 tccctagtat ctttctgtct gcctgtgcgc cccatcagtc gattatcaac aagtaaatct
     1441 gtcttttctt caattaaatc atcaatttca atagctgcta tagggttgcg ttcttcaaga
     1501 taatcataga tatttctacg atcttggcta tgcggtgtga aataccgcac agatgcgtaa
     1561 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     1621 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     1681 gattaagttg ggtaacgcca gggtgattga tgaccgggag ctcacaaaca aggcccagtc
     1741 tttcgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcatggggag
     1801 accccacact accatcggcg ctacggcgtt tcacttctga gttcggcatg gggtcaggtg
     1861 ggaccaccgc gctactgccg ccaggcaaat tctgttttat cagaccgctt ctgcgttctg
     1921 atttaatctg tatcaggctg aaaatcttct ctcatccgcc aaaacagcca agctggagac
     1981 cgtttaaacG CGGCCgcTTA GAACTTAAAT AAGCCTAAGC CATCTTTAAC TTCGTCTAAA
     2041 GTCTGCTGGG TAATGATTCG ACATTTATCA GTACCTTGGC GTAAAACATC CATAATGTAG
     2101 TCAGGATCTT TAGCAAGTTC TTCTCGGCGT TCACGTATAG GTGTAATTAA TTCTTTTAGT
     2161 ACACCTTCAA GACGTTTTTT AACTGTACCA TCACCAAGAC CACCACGGCG GTAGTGTGCT
     2221 TTTAGCTCTT CAACTTCTTC TTTATTTGGA TCAAATGCAT CTAAATAAGT AAATACAATA
     2281 TTTCCTTCAA CTTGACCCGG ATCTTCGATG CGAAGATGAT TTGGATCGGT ATACATCGCA
     2341 TTTACAGCTT TTTTGATGTC TTTGTCAGAA GCATTTAAAA CAATGGTATT GCCTAATGAC
     2401 TTAGACATTT TCGCTTTACC ATCAAAACCC GGTAAACGAG CCATATTAGA AAGCAATGCT
     2461 TTACATTCAG GTAAAAGGTC TTGACCAATT TGACGGTTTA CACGACGCAC GATTTCGTTA
     2521 GTTTGCTCAA TCATTGGAAT TTGATCTTCG CCGCATGGTC CTACAGTCGC TTTAAAAGCC
     2581 GTAATATCAG CTGCTTGTGC TACAGGGTAG CAAAGGAAAC CGGCAGGAAT ATCACGTTCA
     2641 AAACCACGCA TTTGAATTTC AGATTTGATG GTTGGGTTAC GCTCCAAACG AGCTACAGTT
     2701 ACAAAGTTAA GATATAACAT TGTAAGCTCA TTCAGTGCAG GCAAGCATGA CTGTACACAA
     2761 ATTGTGGTTT TAGTTGGATC AATGCCTACC GCTAAATAGT CTAGAGCAAC TTCTAAAATG
     2821 TTACGGCGTA CTTTATCTGG GTTATCGGCA TTGTCTGTTA ACGCTTGTGC ATCTGCTAAT
     2881 AAGAGATGTT GATGATGGCT ATCCTGTAGA CCTACACGAG AACGCAAAGA ACCGACAAAA
     2941 TGCCCGAGGT GAAGTTGTCC GGTTGGGCGG TCGCCTGCCA AAATAATTGG ACGCTGATCA
     3001 ACATTACTCA TgcggccGCa ccTCCTtTGT GAAATTGTTA TCCGCTCACA ATTCCACACA
     3061 TTATACGAGC CGATGATTAA TTGTCAACAG CTCcctaatg caggagtcgc ataagggaga
     3121 gcgtcgagat cccggacacc atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc
     3181 gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg
     3241 cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg
     3301 tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca
     3361 accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca
     3421 gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac
     3481 tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg
     3541 cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg
     3601 accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg
     3661 tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg
     3721 gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa
     3781 gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa
     3841 ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca
     3901 tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg
     3961 cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gacatctcgg
     4021 tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca
     4081 aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg
     4141 gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc
     4201 tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     4261 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag
     4321 ctcactcatt aggcaccggg atctcgaccg atgcccttga gagccttcaa cccagtcagc
     4381 tccttccggt gggcgcgggg catgactaac atgagaatta caacttatat cgtatggggc
     4441 tgacttcagg tgctacattt gaagagataa attgcaagct tatcgatgat aagctgtcaa
     4501 acatgaggcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4561 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4621 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4681 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4741 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4801 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4861 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4921 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4981 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5041 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5101 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5161 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5221 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5281 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5341 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5401 gttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa
     5461 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
     5521 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac
     5581 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga
     5641 ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc
     5701 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt
     5761 gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg
     5821 aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat
     5881 attcttctaa tacctggaat gctgttttcc cagggatcgc agtggtgagt aaccatgcat
     5941 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt
     6001 ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa
     6061 acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga
     6121 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg
     6181 gcctagagca agacgtttcc cgttgaatat ggctcatact cttccttttt caatattatt
     6241 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6301 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     6361 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtc
//