LOCUS       sequence-239475-        6073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A sgRNA expression plasmid for genome editing in Acinetobacter 
            baumannii.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6073)
  AUTHORS   Wang Y, Wang Z, Chen Y, Hua X, Yu Y, Ji Q
  TITLE     A Highly Efficient CRISPR-Cas9-Based Genome Engineering Platform in 
            Acinetobacter baumannii to Understand the H2O2-Sensing Mechanism of 
            OxyR.
  JOURNAL   Cell Chem Biol. 2019 Sep 17. pii: S2451-9456(19)30277-6. doi: 
            10.1016/j.chembiol.2019.09.003.
  PUBMED    31548010
REFERENCE   2  (bases 1 to 6073)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Chem 
            Biol. 2019 Sep 17. pii: S2451-9456(19)30277-6. doi: 
            10.1016/j.chembiol.2019.09.003."
COMMENT     Sequence Label: pSGAb-km
FEATURES             Location/Qualifiers
     source          1..6073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1708..1730
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1722..1739
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1723..1739
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1776..1792
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1777..1793
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(join(1855..3189,3190..3276))
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     promoter        3662..3696
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        3714..3789
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(3822..3838)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3822..3838)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3897..3913)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3897..3913)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3910..3932)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3921..3937
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3945..3951,3952..3969,3970..3975))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3990..4011
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4128..4145)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4299..4887)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4379..4398)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5058..5873)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(5058..5873)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5781..5800
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(5874..5978)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6046..6064
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
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 gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     1741 aattgacgcg tattgggatg gtaccgggcc ccccctcgag gtcgacggta tcgatacggc
     1801 atcagagcag attgtactga gagtgcacca taatcggcat tttcttttgc gtttttattt
     1861 gttaactgtt aattgtcctt gttcaaggat gctgtctttg acaacagatg ttttcttgcc
     1921 tttgatgttc agcaggaagc taggcgcaaa cgttgattgt ttgtctgcgt agaatcctct
     1981 gtttgtcata tagcttgtaa tcacgacatt gtttcctttc gcttgaggta cagcgaagtg
     2041 tgagtaagta aaggttacat cgttaggatc aagatccatt tttaacacaa ggccagtttt
     2101 gttcagcggc ttgtatgggc cagttaaaga attagaaaca taaccaagca tgtaaatatc
     2161 gttagacgta atgccgtcaa tcgtcatttt tgatccgcgg gagtcagtga acagatacca
     2221 tttgccgttc attttaaaga cgttcgcgcg ttcaatttca tctgttactg tgttagatgc
     2281 aatcagcggt ttcatcactt ttttcagtgt gtaatcatcg tttagctcaa tcataccgag
     2341 agcgccgttt gctaactcag ccgtgcgttt tttatcgctt tgcagaagtt tttgactttc
     2401 ttgacggaag aatgatgtgc ttttgccata gtatgctttg ttaaataaag attcttcgcc
     2461 ttggtagcca tcttcagttc cagtgtttgc ttcaaatact aagtatttgt ggcctttatc
     2521 ttctacgtag tgaggatctc tcagcgtatg gttgtcgcct gagctgtagt tgccttcatc
     2581 gatgaactgc tgtacatttt gatacgtttt tccgtcaccg tcaaagattg atttataatc
     2641 ctctacaccg ttgatgttca aagagctgtc tgatgctgat acgttaactt gtgcagttgt
     2701 cagtgtttgt ttgccgtaat gtttaccgga gaaatcagtg tagaataaac ggatttttcc
     2761 gtcagatgta aatgtggctg aacctgacca ttcttgtgtt tggtctttta ggatagaatc
     2821 atttgcatcg aatttgtcgc tgtctttaaa gacgcggcca gcgtttttcc agctgtcaat
     2881 agaagtttcg ccgacttttt gatagaacat gtaaatcgat gtgtcatccg catttttagg
     2941 atctccggct aatgcaaaga cgatgtggta gccgtgatag tttgcgacag tgccgtcagc
     3001 gttttgtaat ggccagctgt cccaaacgtc caggcctttt gcagaagaga tatttttaat
     3061 tgtggacgaa tcgaactcag gaacttgata tttttcattt ttttgctgtt cagggatttg
     3121 cagcatatca tggcgtgtaa tatgggaaat gccgtatgtt tccttatatg gcttttggtt
     3181 cgtttctttc gcaaacgctt gagttgcgcc tcctgccagc agtgcggtag taaaggttaa
     3241 tactgttgct tgttttgcaa actttttgat gttcatcgtt catgtctcct tttttatgta
     3301 ctgtgttagc ggtctgcttc ttccagccct cctgtttgaa gatggcaagt tagttacgca
     3361 caataaaaaa agacctaaaa tatgtaaggg gtgacgccaa agtatacact ttgcccttta
     3421 cacattttag gtcttgcctg ctttatcagt aacaaacccg cgcgatttac ttttcgacct
     3481 cattctatta gactctcgtt tggattgcaa ctggtctatt ttcctctttt gtttgataga
     3541 aaatcataaa aggatttgca gactacgggc ctaaagaact aaaaaatcta tctgtttctt
     3601 ttcattctct gtatttttta tagtttctgt tgcatgggca taaagttgca agcttgatat
     3661 cttgacagct agctcagtcc taggtataat actagtcgag accattggtc tcagttttag
     3721 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
     3781 agtcggtgct ttttttgata tcgaattcct gcagcccggg ggatccacta gttctagagc
     3841 ggccgccacc gcggtggagc tcatcccaat ggcgcgccga gcttggctcg agcatggtca
     3901 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     3961 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4021 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4081 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4141 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4201 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4261 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4321 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4381 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4441 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4501 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4561 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4621 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4681 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4741 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4801 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4861 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4921 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4981 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5041 taaacttggt ctgacagtta gaaaaactca tcgagcatca aatgaaactg caatttattc
     5101 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
     5161 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
     5221 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     5281 tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag
     5341 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     5401 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa
     5461 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     5521 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccagg gatcgcagtg
     5581 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     5641 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     5701 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc
     5761 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     5821 ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct catactcttc
     5881 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5941 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6001 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     6061 aggccctttc gtc
//
