LOCUS       sequence-379791-        8109 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 8109)
  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 8109)
  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-A
FEATURES             Location/Qualifiers
     source          1..8109
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    135..156
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(171..176,177..194,195..201)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    209..225
                     /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             243..254
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             261..938
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      983..1054
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1070..1097
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      5064..5121
                     /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(5145..5162)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5316..5904)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5396..5415)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(6087..6181)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      complement(6206..6292)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(6310..7104)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     CDS             complement(7200..7859)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7774..7793
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     8021..8039
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(8043..8086)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    join(8089..8109,1)
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 ggtctcacta tatctagaga cgtctagaca tatgcaatac gcaaaccgcc tctccccgcg
       61 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
      121 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
      181 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacatactag
      241 agaaagagga gaaatactag atggcttcct ccgaagacgt tatcaaagag ttcatgcgtt
      301 tcaaagttcg tatggaaggt tccgttaacg gtcacgagtt cgaaatcgaa ggtgaaggtg
      361 aaggtcgtcc gtacgaaggt acccagaccg ctaaactgaa agttaccaaa ggtggtccgc
      421 tgccgttcgc ttgggacatc ctgtccccgc agttccagta cggttccaaa gcttacgtta
      481 aacacccggc tgacatcccg gactacctga aactgtcctt cccggaaggt ttcaaatggg
      541 aacgtgttat gaacttcgaa gacggtggtg ttgttaccgt tacccaggac tcctccctgc
      601 aagacggtga gttcatctac aaagttaaac tgcgtggtac caacttcccg tccgacggtc
      661 cggttatgca gaaaaaaacc atgggttggg aagcttccac cgaacgtatg tacccggaag
      721 acggtgctct gaaaggtgaa atcaaaatgc gtctgaaact gaaagacggt ggtcactacg
      781 acgctgaagt taaaaccacc tacatggcta aaaaaccggt tcagctgccg ggtgcttaca
      841 aaaccgacat caaactggac atcacctccc acaacgaaga ctacaccatc gttgaacagt
      901 acgaacgtgc tgaaggtcgt cactccaccg gtgcttaata acgctgatag tgctagtgta
      961 gatcgctact agagccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     1021 tcgttttatc tgttgtttgt cggtgaacgc tctctactag agtcacactg gctcaccttc
     1081 gggtgggcct ttctgcgttt atagcatgcc gtctcattag gtacagagac cctgcagttg
     1141 caagacaata tcgaccgctt gcagcttgat aaagtgatca aagagggacg agcagctaca
     1201 gcactgctaa gggagatcgg agagccactg ccgccaccag tgaaagtgaa aacaggttgg
     1261 tttagtgctg attattacgg ctttgaggac atgctcacac gttgggataa tgatcaatct
     1321 agcctttcca tttatcacag agagataaaa gaggagcaac aggaacagca acgaaaacaa
     1381 cagcagatag aaagagaaac taaacaaaaa caggaagatg acaaaaagaa acggcaggac
     1441 tatgaaagat atttatcaaa atacggctat gtcaaagatg ctgagtatga ccatatcagc
     1501 gaccatgttg cttcagacat ttccatgttt acacgtcttc aggcgcaggc atgggcaagt
     1561 aacgatatgc aagagtacat aaggtgttca aagcaaaaat acgagcagat cagcgacaga
     1621 atagcctatg agcgtgactt gaaaaaaatc gggcaattga gccgtatcct ggataaagac
     1681 agacaggcac ttgctgatat tttaccgcag ttgatgcctt actatgagca gatgcaacag
     1741 gctgttaata aacggaaaat tttacttgag aatgagcgac agccatcatt taaaccacgt
     1801 tatgaccaag aacagcctaa gccgaaaaaa gacaatgacc taacttttta atgtgttaca
     1861 atagcattca aatgtaacac attggagtga aaaacatgac agaagcgact tttacctttc
     1921 gggtagacca tgacctaaag caagaatttt caagccttgc taagactgtt gatcggtcgg
     1981 gggcgcagct aatccgtgac ttcatgcgtg actttgttaa gaagcaacag gaagccgcag
     2041 actatgacaa atggttcaag cagcaggtac agatcggctt aaatgaagcc aatgcaggca
     2101 agttgattcc acatgaggac gtaaaggcag agtttgccgc tagacgtgct gcaacattgg
     2161 cgaaactggc agcgaagaat gaagattgag tggactgaaa cagctcgcca agatcgtaga
     2221 aatatctatg attatcttga agaacgcaac cctatagcag ctattgaaat tgatgattta
     2281 attgaagaaa agacagattt acttgttgat aatcgactga tggggcgcac aggcagacag
     2341 aaagatacta gggagttagt gatacatccg cattatgtgg ttgtatatga catcactgat
     2401 ataatacgga tactcagagt gctacacaca tcgcaggagt ggtcatgact tactcatgta
     2461 ctttggatta tttagtgtta taaaatcctg atttataaat tttttttgtt aaaaaagata
     2521 aaagcccctt gcaattgctt ggggctttac cgtaatttat ggggtacaga tcttcgatac
     2581 tgacatatcg gcaatcgaaa gcattaaggt ttgacgaccg ctaatgattt caccacaggg
     2641 gcttaatgta cctgtcttaa attctaaggt tttaactcgc tttgtcaagc atagacccca
     2701 aaaatttagc caatgtctgt aactcaatct gtccatgtgt gggtgatgag gtacagtgac
     2761 gctagcacac atcggaaaaa cgctattact aggggaactg aacagagtag cggacgcaat
     2821 gagtagtcat ttaattggcg gttatgagcg tgttcaggcg gtgctatcaa tcgtaatcat
     2881 aacagtggca gcttgataca gtgatgtcat ccctgatgcg aaagcgaccg accgacggta
     2941 catcgaatgg gaatacttta gggtgatttt taagaatcgc tctagggtga gtatttccca
     3001 ttcagctctg ctccctccct ctggtacttt aatcaaaagc actactaaac atatgttttt
     3061 aaataaaaaa tattgatata gagataatat tagtaagaat aattaaacaa ttgaatatag
     3121 ataaatcatt gttaaataaa gattaattat taaaatgaat gtatacttat atataaatca
     3181 atgatttaaa atatttgata aagaaaactt ttcaaaaaaa atataattga gattgtgtca
     3241 tttcggtcaa ttcttaatat gttccacgca agttttagct atggtgctaa acagaaattt
     3301 gctgaaaaag aacttttcac tgaactggtt aaaatgtaag cagcctgaga gccgccaaaa
     3361 attttaaaaa caaaccgcct taatcatctt caaaaaatac ctctaaaacc tcaccatttg
     3421 cgttttaaga cccatatttc atcctgccct tatgttccca tgctgatagc tataaagtgt
     3481 ctgtaatcgc ttcctatgac gttctaggct gttgataact tttggaacaa cgcaaaatgt
     3541 taaaatccga tcatttttta acctagttat tttcgttaca gggtaacttt ggtagtatta
     3601 tttcaatatt tagttgttac aggataacaa aatatatgtt acagggtaat taatttagcg
     3661 ttacaggata actatagtta cagggtaacg ttaaatagtt atcctgtaat ttaaaaatag
     3721 ttgcaggata gcataaattt ttacaaataa attttataaa aagttatcct gtaactaaaa
     3781 acaatgttat cctgtaacta tgatatacta tatgggagtt aaagacatga aagacccgaa
     3841 cgatcagaaa acacaggaca tgctaaaaga gccgccaaaa accaatgccg aacgacaaaa
     3901 agcatatcgt gaaaaacgca aaagccttga tagtcaacgt ttggaggtgt ttatagataa
     3961 gggtgtgtcg gatatgcttg cggacatggt gggcgcagca ggggagagcc aaaaagccat
     4021 tttgaccgca ttgattgaga aagagtataa gcggctgtat gcagtgaaaa atagcaagct
     4081 aaaaaagtac attgattcaa aaagtaatgc agataaaaaa gaaaccctcg atttatgagg
     4141 gttttttata actactaatg cctatgaaaa taagtgctta tcattttctc taatgtaccc
     4201 aataaaatcg gggttttgtt ccacaagttt cttgatgtca ttatcatgtt tcttcagtaa
     4261 gtaagccccg atcaggatct tgcgttttgc atcgtctgac cttgcctgtt tttgctctcg
     4321 acctttcacc acagcgtcag ctagatcctt tcttgccttt tcctgacgtt ccaatttttc
     4381 cttttcttca tcgagcattt gacccaattc catagctgtt tttttgacca tttttaatgc
     4441 ctgtgtctca acctattttc cgaagtatga accatcatca atgtgaaaag tgttctactt
     4501 caaaaactcg acttacatca agatccgagc gcagcgagca aaatcaaaaa acaaagtcaa
     4561 agccttattg ctcttgcttt tgcttttagc ctcgcagagt tcccgaaggg cgcacttacg
     4621 caaaattttt gctacgccaa attttgcaag tacggtcagg gaaaccccga caccccaacc
     4681 gcccaaaact tggggcggct gttaataaac aggtgatgaa aatggcaatt taccattgtg
     4741 aaatgcagaa catttcgagg tcagatggtc gctcaatcgt ggcatgtgca gcataccgag
     4801 caggcgaaaa attgtactgt gatacgtacg gaaaagagca ggactacacc aaaaaaacag
     4861 gcattgaata cacccaaatt tttgccccaa ctggggcaag tcctgacatg ttagatcgtc
     4921 aaaccctatg gaatcgggta gagcaatccg aactaaaaaa gaacggtgac atcaaacagg
     4981 aggcaagatt agcaaaggaa gtagagatag cattgccgca tgaactggat aagacacagc
     5041 gtcaggcaac ttgttaactg cagtccggca aaaaagggca aggtgtcacc accctgccct
     5101 ttttctttaa aaccgaaaag attacttcgc gttatgcagg cttcctcgct cactgactcg
     5161 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5221 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5281 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc acaggctccg cccccctgac
     5341 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5401 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5461 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5521 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5581 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5641 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5701 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5761 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5821 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5881 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5941 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6001 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6061 acttggtctg acagctcgag gcttggattc tcaccaataa aaaacgcccg gcggcaaccg
     6121 agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt
     6181 ccaagcgagc tcgatattct cgaggatttg tcctactcag gagagcgttc accgacaaac
     6241 aacagataaa acgaaaggcc cagtctttcg actgagcctt tcgttttatt tgatgccgcc
     6301 ggcgacgtcc taaaacaatt catccagtaa aatataatat tttattttct cccaatcagg
     6361 cttgatcccc agtaagtcaa aaaatagctc gacatactgt tcttccccga tatcctccct
     6421 gatcgaccgg acgcagaagg caatgtcata ccacttgtcc gccctgccgc ttctcccaag
     6481 atcaataaag ccacttactt tgccatcttt cacaaagatg ttgctgtctc ccaggtcgcc
     6541 gtgggaaaag acaagttcct cttcgggctt ttccgtcttt aaaaaatcat acagctcgcg
     6601 cggatcttta aatggagtgt cttcttccca gttttcgcaa tccacatcgg ccagatcgtt
     6661 attcagtaag taatccaatt cggctaagcg gctgtctaag ctattcgtat agggacaatc
     6721 cgatatgtcg atggagtgaa agagcctgat gcactccgca tacagctcga taatcttttc
     6781 agggctttgt tcatcttcat actcttccga gcaaaggacg ccatcggcct cactcatgag
     6841 cagattgctc cagccatcat gccgttcaaa gtgcaggacc tttggaacag gcagctttcc
     6901 ttccagccat agcatcatgt ccttttcccg ttccacatca taggtggtcc ctttataccg
     6961 gctgtccgtc atttttaaat ataggttttc attttctccc accagcttat ataccttagc
     7021 aggagacatt ccttccgtat cttttacgca gcggtatttt tcgatcagtt ttttcaattc
     7081 cggtgatatt ctcattttag ccatttatta tttccttcct cttttctaca gtatttaaag
     7141 attatatcac attatccatt aaaaatcaaa cggatcccca gcttctcgag aatatcaaat
     7201 tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca
     7261 tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg
     7321 ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc
     7381 acgtttaaat caaaactggt gaaactcacc cagggattgg ctgacacgaa aaacatattc
     7441 tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa
     7501 tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt
     7561 tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca
     7621 ccgtctttca ttgccatacg aaattccgga tgagcattca tcaggcgggc aagaatgtga
     7681 ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata
     7741 tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt
     7801 tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt
     7861 ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt
     7921 atttcattat ggtgaaagtt ggaacctctt acgtgcccga tcaactcgag tgccacctga
     7981 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggca
     8041 gaatttcaga taaaaaaaat ccttagcttt cgctaaggat gatttctgga attcgcggcc
     8101 gcttctaga
//
