LOCUS       sequence-379792-        8005 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 2 MoClo plasmid for A. baumannii.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8005)
  AUTHORS   Brychcy M, Kokodynski A, Lloyd D, Godoy VG
  TITLE     AspFlex: Molecular Tools to Study Gene Expression and Regulation in 
            Acinetobacter baumannii.
  JOURNAL   ACS Synth Biol. 2023 Aug 16. doi: 10.1021/acssynbio.3c00167.
  PUBMED    37587063
REFERENCE   2  (bases 1 to 8005)
  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: "ACS Synth 
            Biol. 2023 Aug 16. doi: 10.1021/acssynbio.3c00167."
COMMENT     Sequence Label: pMB2-B
FEATURES             Location/Qualifiers
     source          1..8005
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    123..144
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(159..164,165..182,183..189)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    197..213
                     /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)."
     RBS             231..242
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             249..926
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      971..1042
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1058..1085
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      5046..5103
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(5127..5144)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5298..5886)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5378..5397)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(6069..6163)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             6182..6976
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     terminator      6994..7080
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(7096..7755)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7670..7689
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     7917..7935
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(7939..7982)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    join(7985..8005,1)
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 ggtctcagta catctagaga cgcaatacgc aaaccgcctc tccccgcgcg ttggccgatt
       61 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
      121 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
      181 cgtatgttgt gtggaattgt gagcggataa caatttcaca catactagag aaagaggaga
      241 aatactagat ggcttcctcc gaagacgtta tcaaagagtt catgcgtttc aaagttcgta
      301 tggaaggttc cgttaacggt cacgagttcg aaatcgaagg tgaaggtgaa ggtcgtccgt
      361 acgaaggtac ccagaccgct aaactgaaag ttaccaaagg tggtccgctg ccgttcgctt
      421 gggacatcct gtccccgcag ttccagtacg gttccaaagc ttacgttaaa cacccggctg
      481 acatcccgga ctacctgaaa ctgtccttcc cggaaggttt caaatgggaa cgtgttatga
      541 acttcgaaga cggtggtgtt gttaccgtta cccaggactc ctccctgcaa gacggtgagt
      601 tcatctacaa agttaaactg cgtggtacca acttcccgtc cgacggtccg gttatgcaga
      661 aaaaaaccat gggttgggaa gcttccaccg aacgtatgta cccggaagac ggtgctctga
      721 aaggtgaaat caaaatgcgt ctgaaactga aagacggtgg tcactacgac gctgaagtta
      781 aaaccaccta catggctaaa aaaccggttc agctgccggg tgcttacaaa accgacatca
      841 aactggacat cacctcccac aacgaagact acaccatcgt tgaacagtac gaacgtgctg
      901 aaggtcgtca ctccaccggt gcttaataac gctgatagtg ctagtgtaga tcgctactag
      961 agccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     1021 ttgtttgtcg gtgaacgctc tctactagag tcacactggc tcaccttcgg gtgggccttt
     1081 ctgcgtttat acgtctcatt agggacagag accctgcagt taacaagttg cctgacgctg
     1141 tgtcttatcc agttcatgcg gcaatgctat ctctacttcc tttgctaatc ttgcctcctg
     1201 tttgatgtca ccgttctttt ttagttcgga ttgctctacc cgattccata gggtttgacg
     1261 atctaacatg tcaggacttg ccccagttgg ggcaaaaatt tgggtgtatt caatgcctgt
     1321 ttttttggtg tagtcctgct cttttccgta cgtatcacag tacaattttt cgcctgctcg
     1381 gtatgctgca catgccacga ttgagcgacc atctgacctc gaaatgttct gcatttcaca
     1441 atggtaaatt gccattttca tcacctgttt attaacagcc gccccaagtt ttgggcggtt
     1501 ggggtgtcgg ggtttccctg accgtacttg caaaatttgg cgtagcaaaa attttgcgta
     1561 agtgcgccct tcgggaactc tgcgaggcta aaagcaaaag caagagcaat aaggctttga
     1621 ctttgttttt tgattttgct cgctgcgctc ggatcttgat gtaagtcgag tttttgaagt
     1681 agaacacttt tcacattgat gatggttcat acttcggaaa ataggttgag acacaggcat
     1741 taaaaatggt caaaaaaaca gctatggaat tgggtcaaat gctcgatgaa gaaaaggaaa
     1801 aattggaacg tcaggaaaag gcaagaaagg atctagctga cgctgtggtg aaaggtcgag
     1861 agcaaaaaca ggcaaggtca gacgatgcaa aacgcaagat cctgatcggg gcttacttac
     1921 tgaagaaaca tgataatgac atcaagaaac ttgtggaaca aaaccccgat tttattgggt
     1981 acattagaga aaatgataag cacttatttt cataggcatt agtagttata aaaaaccctc
     2041 ataaatcgag ggtttctttt ttatctgcat tactttttga atcaatgtac ttttttagct
     2101 tgctattttt cactgcatac agccgcttat actctttctc aatcaatgcg gtcaaaatgg
     2161 ctttttggct ctcccctgct gcgcccacca tgtccgcaag catatccgac acacccttat
     2221 ctataaacac ctccaaacgt tgactatcaa ggcttttgcg tttttcacga tatgcttttt
     2281 gtcgttcggc attggttttt ggcggctctt ttagcatgtc ctgtgttttc tgatcgttcg
     2341 ggtctttcat gtctttaact cccatatagt atatcatagt tacaggataa cattgttttt
     2401 agttacagga taacttttta taaaatttat ttgtaaaaat ttatgctatc ctgcaactat
     2461 ttttaaatta caggataact atttaacgtt accctgtaac tatagttatc ctgtaacgct
     2521 aaattaatta ccctgtaaca tatattttgt tatcctgtaa caactaaata ttgaaataat
     2581 actaccaaag ttaccctgta acgaaaataa ctaggttaaa aaatgatcgg attttaacat
     2641 tttgcgttgt tccaaaagtt atcaacagcc tagaacgtca taggaagcga ttacagacac
     2701 tttatagcta tcagcatggg aacataaggg caggatgaaa tatgggtctt aaaacgcaaa
     2761 tggtgaggtt ttagaggtat tttttgaaga tgattaaggc ggtttgtttt taaaattttt
     2821 ggcggctctc aggctgctta cattttaacc agttcagtga aaagttcttt ttcagcaaat
     2881 ttctgtttag caccatagct aaaacttgcg tggaacatat taagaattga ccgaaatgac
     2941 acaatctcaa ttatattttt tttgaaaagt tttctttatc aaatatttta aatcattgat
     3001 ttatatataa gtatacattc attttaataa ttaatcttta tttaacaatg atttatctat
     3061 attcaattgt ttaattattc ttactaatat tatctctata tcaatatttt ttatttaaaa
     3121 acatatgttt agtagtgctt ttgattaaag taccagaggg agggagcaga gctgaatggg
     3181 aaatactcac cctagagcga ttcttaaaaa tcaccctaaa gtattcccat tcgatgtacc
     3241 gtcggtcggt cgctttcgca tcagggatga catcactgta tcaagctgcc actgttatga
     3301 ttacgattga tagcaccgcc tgaacacgct cataaccgcc aattaaatga ctactcattg
     3361 cgtccgctac tctgttcagt tcccctagta atagcgtttt tccgatgtgt gctagcgtca
     3421 ctgtacctca tcacccacac atggacagat tgagttacag acattggcta aatttttggg
     3481 gtctatgctt gacaaagcga gttaaaacct tagaatttaa gacaggtaca ttaagcccct
     3541 gtggtgaaat cattagcggt cgtcaaacct taatgctttc gattgccgat atgtcagtat
     3601 cgaagatctg taccccataa attacggtaa agccccaagc aattgcaagg ggcttttatc
     3661 ttttttaaca aaaaaaattt ataaatcagg attttataac actaaataat ccaaagtaca
     3721 tgagtaagtc atgaccactc ctgcgatgtg tgtagcactc tgagtatccg tattatatca
     3781 gtgatgtcat atacaaccac ataatgcgga tgtatcacta actccctagt atctttctgt
     3841 ctgcctgtgc gccccatcag tcgattatca acaagtaaat ctgtcttttc ttcaattaaa
     3901 tcatcaattt caatagctgc tatagggttg cgttcttcaa gataatcata gatatttcta
     3961 cgatcttggc gagctgtttc agtccactca atcttcattc ttcgctgcca gtttcgccaa
     4021 tgttgcagca cgtctagcgg caaactctgc ctttacgtcc tcatgtggaa tcaacttgcc
     4081 tgcattggct tcatttaagc cgatctgtac ctgctgcttg aaccatttgt catagtctgc
     4141 ggcttcctgt tgcttcttaa caaagtcacg catgaagtca cggattagct gcgcccccga
     4201 ccgatcaaca gtcttagcaa ggcttgaaaa ttcttgcttt aggtcatggt ctacccgaaa
     4261 ggtaaaagtc gcttctgtca tgtttttcac tccaatgtgt tacatttgaa tgctattgta
     4321 acacattaaa aagttaggtc attgtctttt ttcggcttag gctgttcttg gtcataacgt
     4381 ggtttaaatg atggctgtcg ctcattctca agtaaaattt tccgtttatt aacagcctgt
     4441 tgcatctgct catagtaagg catcaactgc ggtaaaatat cagcaagtgc ctgtctgtct
     4501 ttatccagga tacggctcaa ttgcccgatt tttttcaagt cacgctcata ggctattctg
     4561 tcgctgatct gctcgtattt ttgctttgaa caccttatgt actcttgcat atcgttactt
     4621 gcccatgcct gcgcctgaag acgtgtaaac atggaaatgt ctgaagcaac atggtcgctg
     4681 atatggtcat actcagcatc tttgacatag ccgtattttg ataaatatct ttcatagtcc
     4741 tgccgtttct ttttgtcatc ttcctgtttt tgtttagttt ctctttctat ctgctgttgt
     4801 tttcgttgct gttcctgttg ctcctctttt atctctctgt gataaatgga aaggctagat
     4861 tgatcattat cccaacgtgt gagcatgtcc tcaaagccgt aataatcagc actaaaccaa
     4921 cctgttttca ctttcactgg tggcggcagt ggctctccga tctcccttag cagtgctgta
     4981 gctgctcgtc cctctttgat cactttatca agctgcaagc ggtcgatatt gtcttgcaac
     5041 tgcagtccgg caaaaaaggg caaggtgtca ccaccctgcc ctttttcttt aaaaccgaaa
     5101 agattacttc gcgttatgca ggcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5161 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5221 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5281 gccgcgttgc tggcgttttt ccacaggctc cgcccccctg acgagcatca caaaaatcga
     5341 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5401 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5461 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5521 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5581 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5641 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5701 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5761 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5821 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5881 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5941 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6001 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagctcg
     6061 aggcttggat tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc
     6121 agatggagtt ctgaggtcat tactggatct atcaacagga gtccaagcga gctcgatata
     6181 aatggctaaa atgagaatat caccggaatt gaaaaaactg atcgaaaaat accgctgcgt
     6241 aaaagatacg gaaggaatgt ctcctgctaa ggtatataag ctggtgggag aaaatgaaaa
     6301 cctatattta aaaatgacgg acagccggta taaagggacc acctatgatg tggaacggga
     6361 aaaggacatg atgctatggc tggaaggaaa gctgcctgtt ccaaaggtcc tgcactttga
     6421 acggcatgat ggctggagca atctgctcat gagtgaggcc gatggcgtcc tttgctcgga
     6481 agagtatgaa gatgaacaaa gccctgaaaa gattatcgag ctgtatgcgg agtgcatcag
     6541 gctctttcac tccatcgaca tatcggattg tccctatacg aatagcttag acagccgctt
     6601 agccgaattg gattacttac tgaataacga tctggccgat gtggattgcg aaaactggga
     6661 agaagacact ccatttaaag atccgcgcga gctgtatgat tttttaaaga cggaaaagcc
     6721 cgaagaggaa cttgtctttt cccacggcga cctgggagac agcaacatct ttgtgaaaga
     6781 tggcaaagta agtggcttta ttgatcttgg gagaagcggc agggcggaca agtggtatga
     6841 cattgccttc tgcgtccggt cgatcaggga ggatatcggg gaagaacagt atgtcgagct
     6901 attttttgac ttactgggga tcaagcctga ttgggagaaa ataaaatatt atattttact
     6961 ggatgaattg ttttaggacg tcgccggcgg catcaaataa aacgaaaggc tcagtcgaaa
     7021 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     7081 cctcgagaaa tcaaattacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt
     7141 aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg
     7201 catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa
     7261 gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga
     7321 cacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca
     7381 cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca
     7441 gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc
     7501 ccatatcacc agctcaccgt ctttcattgc catacgaaat tccggatgag cattcatcag
     7561 gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt
     7621 taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg
     7681 aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt
     7741 gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac
     7801 gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt gcccgatcaa
     7861 ctcgagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     7921 ggcgtatcac gaggcagaat ttcagataaa aaaaatcctt agctttcgct aaggatgatt
     7981 tctggaattc gcggccgctt ctaga
//
