LOCUS       sequence-404862-       10260 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 MoClo plasmid for A. baumannii with additional Tellurite 
            resistance cassette for multi-drug resistance strains.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10260)
  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 10260)
  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: pMB1-A-TelR
FEATURES             Location/Qualifiers
     source          1..10260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             2815..2837
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    3085..3106
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3121..3126,3127..3144,3145..3151)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3159..3175
                     /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             3193..3204
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3211..3888
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      3933..4004
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4020..4047
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(8013..8030)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8184..8772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8264..8283)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8943..9734,9735..9803))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     9566..9585
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9804..9908)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9976..9994
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(9998..10041)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    10044..10063
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     terminator      10143..10174
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 tgccagccca acaaagccgg ttgccgccgc cgccaggatc gcgccgatga tgccggccac
       61 accggccatc gcccaccagg tcgccgcctt ccggttccat tcctgctggt actgcttcgc
      121 aatgctggac ctcggctcac cataggctga ccgctcgatg gcgtatgccg cttctcccct
      181 tggcgtaaaa cccagcgccg caggcggcat tgccatgctg cccgccgctt tcccgaccac
      241 gacgcgcgca ccaggcttgc ggtccagacc ttcggccacg gcgagctgcg caaggacata
      301 atcagccgcc gacttggctc cacgcgcctc gatcagctct tgcactcgcg cgaaatcctt
      361 ggcctccacg gccgccatga atcgcgcacg cggcgaaggc tccgcagggc cggcgtcgtg
      421 atcgccgccg agaatgccct tcaccaagtt cgacgacacg aaaatcatgc tgacggctat
      481 caccatcatg cagacggatc gcacgaaccc gcagaactca cccccgaaca cgagcacggc
      541 acccgcgacc actatgccaa gaatgcccaa ggtaaaaatt gccggccccg ccatgaagtc
      601 cgtgaatgcc ccgacggccg aagtgaaggg caggccgcca cccaggccgc cgccctcact
      661 gcccggcacc tggtcgctga atgtcgatgc cagcacctgc ggcacgtcaa tgcttccggg
      721 cgtcgcgctc gggctgatcg cccatcccgt tactgccccg atcccggcaa tggcaaggac
      781 tgccagcgcc gcgatgagga agcgggtgcc ccgcttcttc atcttcgcgc ctcgggcctc
      841 caggccgcct acctgggcga aaacatcggt gtttgtggca ttcatacgga ctcctgttgg
      901 gccagctcgc gcacgggctg gcgggtcagc ttggcttgaa gatcgccacg cattgcggcg
      961 atctgcttct cggcatcctt gcgcttctgc acgccttcct gctggatgcg aataacgtcc
     1021 tcgacggtct tgatgagcgt cgtctgaacc tgcttgagcg tgtccacgtc gatcaccagg
     1081 cgttggttct ccttcgccgt ttcgacggac gtgcgatgca gcagggccgc attgcgcttc
     1141 atcaggtcgt tggtggtgtc gtcgatggcc gtggccagtt cgacggcgtt cttctgctcg
     1201 ttgaggctca aggccagcat gaattgccgc ttccacgccg gcacggtgat ttcgcggatg
     1261 gtgtggaatt tatcgaccag catctggttg ttggcctgga tcatgcggat ggtcggcagg
     1321 ctctgcatgg ccgaatgttg caaggcgatc aggtcgccga tgcgcttgtc caggttggca
     1381 accatcgcat cgaggtcggc cagctcctgc acgcggccag ggtcgttccc gacattgccg
     1441 cgcagaccct cggcctgctc gcgcagctcg gcaaggcgga ccttgccggc cgcgatgtgg
     1501 acgccaagaa ggcggtgttc ctcgcgcacg gctgcgaaca tttcgtcgag cgaggcattg
     1561 cgctgcgcga tgccttgctg ggtggtctgc acttcgctga ccaggtgttc gatctgctcg
     1621 cgggtcgtgt cgaagcgcgc catgaagccc gtcgaacgga cgcgggagcg gtcgatcagc
     1681 gggccaatca ggggcaggcg ggaacggttg tcggacaaag ggccgacgtt cagggaacgg
     1741 gccttggcga caacctgggt cagtttctcg cctgcttcgt ccaggtcgct gttgcgcacc
     1801 tggtccagca ggctatcggc gtagcgggac gtgtgctcgg ccacgtcgcg gccgaactcg
     1861 gcaacggtct gcggactgcc gacctcgatc cgctgcgcga ccgcatggac ttccggcacg
     1921 tcgctttcct gcaagcccag ctcgcgcagg gttgccgggg tcatgtcaaa ggcgacgata
     1981 ggggccttgg cgtcgtgcgt cgttttcagt gcgttcatag ggttctcccg ccgtgttatt
     2041 ggttgatgcc ttccaggctc tgcgaaaggc tccgcatgag cgcctggtga gctttggccg
     2101 cctcggcgac cattgccgga ttcatgttct tggtggtgat gagcgcgagg gtgtgctgac
     2161 gccagacggg caccaggacg gatgccgttt cagagaagcg gtccagcatg tccacggcct
     2221 gcgcccgcgt gagcttcatc tgagtgacgc tcatttcatg ggacgccatg agggttgcca
     2281 ggttggcgag cttgcgcgcg aagcgttcgc gcggcttgtc gaactcgatc acgccggcct
     2341 tggccgcgcc ggcctcgggg ttctcgtcca ggaactcgcg cccggcttga atgtaggctc
     2401 tgagccggtc tacctcggcc tcatgcgtat tgagcatgtc atccaaggcg cgcaacgtgt
     2461 cccgcacgcg ctgcgctacg ccctcggctt cgtccagcaa ctggtcgagc gtcttgcggg
     2521 cgacctgata cctcacctgg cgttcaacct cacggccaag catcttctcg aaccaggtag
     2581 gcttttccgc gatcttgcgg gggtccgcgt cgaccagctt cgccacgatc tggctgattt
     2641 tgtcggccag cgcggcaact gcgccgtgct ccatcagatt cgacagctcg ttgagggaat
     2701 ccgccccgtc gatgccggcc ccgtactcgc caatcgtcgc cggcgacgcg aagagggcgg
     2761 gcaaaacctc ccccttcaat cgcgccatgt tcacgcttgt tcttccatgg tatatctcct
     2821 tcttaaagtt aaacaaaatt attcggaacc cagcatgata ttccggaaat accaactaag
     2881 tcaacggctg atatccattg ctgttgacaa agggaatcag gggatcttga agttcctatt
     2941 ccgaagttcc tattctctag aacgtctcaa tctctataga gacccaatac gcaaaccgcc
     3001 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     3061 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     3121 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     3181 cacatactag agaaagagga gaaatactag atggcttcct ccgaagacgt tatcaaagag
     3241 ttcatgcgtt tcaaagttcg tatggaaggt tccgttaacg gtcacgagtt cgaaatcgaa
     3301 ggtgaaggtg aaggtcgtcc gtacgaaggt acccagaccg ctaaactgaa agttaccaaa
     3361 ggtggtccgc tgccgttcgc ttgggacatc ctgtccccgc agttccagta cggttccaaa
     3421 gcttacgtta aacacccggc tgacatcccg gactacctga aactgtcctt cccggaaggt
     3481 ttcaaatggg aacgtgttat gaacttcgaa gacggtggtg ttgttaccgt tacccaggac
     3541 tcctccctgc aagacggtga gttcatctac aaagttaaac tgcgtggtac caacttcccg
     3601 tccgacggtc cggttatgca gaaaaaaacc atgggttggg aagcttccac cgaacgtatg
     3661 tacccggaag acggtgctct gaaaggtgaa atcaaaatgc gtctgaaact gaaagacggt
     3721 ggtcactacg acgctgaagt taaaaccacc tacatggcta aaaaaccggt tcagctgccg
     3781 ggtgcttaca aaaccgacat caaactggac atcacctccc acaacgaaga ctacaccatc
     3841 gttgaacagt acgaacgtgc tgaaggtcgt cactccaccg gtgcttaata acgctgatag
     3901 tgctagtgta gatcgctact agagccaggc atcaaataaa acgaaaggct cagtcgaaag
     3961 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag agtcacactg
     4021 gctcaccttc gggtgggcct ttctgcgttt ataggtctca tgtttgccag agacgctgca
     4081 gttaacaagt tgcctgacgc tgtgtcttat ccagttcatg cggcaatgct atctctactt
     4141 cctttgctaa tcttgcctcc tgtttgatgt caccgttctt ttttagttcg gattgctcta
     4201 cccgattcca tagggtttga cgatctaaca tgtcaggact tgccccagtt ggggcaaaaa
     4261 tttgggtgta ttcaatgcct gtttttttgg tgtagtcctg ctcttttccg tacgtatcac
     4321 agtacaattt ttcgcctgct cggtatgctg cacatgccac gattgagcga ccatctgacc
     4381 tcgaaatgtt ctgcatttca caatggtaaa ttgccatttt catcacctgt ttattaacag
     4441 ccgccccaag ttttgggcgg ttggggtgtc ggggtttccc tgaccgtact tgcaaaattt
     4501 ggcgtagcaa aaattttgcg taagtgcgcc cttcgggaac tctgcgaggc taaaagcaaa
     4561 agcaagagca ataaggcttt gactttgttt tttgattttg ctcgctgcgc tcggatcttg
     4621 atgtaagtcg agtttttgaa gtagaacact tttcacattg atgatggttc atacttcgga
     4681 aaataggttg agacacaggc attaaaaatg gtcaaaaaaa cagctatgga attgggtcaa
     4741 atgctcgatg aagaaaagga aaaattggaa cgtcaggaaa aggcaagaaa ggatctagct
     4801 gacgctgtgg tgaaaggtcg agagcaaaaa caggcaaggt cagacgatgc aaaacgcaag
     4861 atcctgatcg gggcttactt actgaagaaa catgataatg acatcaagaa acttgtggaa
     4921 caaaaccccg attttattgg gtacattaga gaaaatgata agcacttatt ttcataggca
     4981 ttagtagtta taaaaaaccc tcataaatcg agggtttctt ttttatctgc attacttttt
     5041 gaatcaatgt acttttttag cttgctattt ttcactgcat acagccgctt atactctttc
     5101 tcaatcaatg cggtcaaaat ggctttttgg ctctcccctg ctgcgcccac catgtccgca
     5161 agcatatccg acacaccctt atctataaac acctccaaac gttgactatc aaggcttttg
     5221 cgtttttcac gatatgcttt ttgtcgttcg gcattggttt ttggcggctc ttttagcatg
     5281 tcctgtgttt tctgatcgtt cgggtctttc atgtctttaa ctcccatata gtatatcata
     5341 gttacaggat aacattgttt ttagttacag gataactttt tataaaattt atttgtaaaa
     5401 atttatgcta tcctgcaact atttttaaat tacaggataa ctatttaacg ttaccctgta
     5461 actatagtta tcctgtaacg ctaaattaat taccctgtaa catatatttt gttatcctgt
     5521 aacaactaaa tattgaaata atactaccaa agttaccctg taacgaaaat aactaggtta
     5581 aaaaatgatc ggattttaac attttgcgtt gttccaaaag ttatcaacag cctagaacgt
     5641 cataggaagc gattacagac actttatagc tatcagcatg ggaacataag ggcaggatga
     5701 aatatgggtc ttaaaacgca aatggtgagg ttttagaggt attttttgaa gatgattaag
     5761 gcggtttgtt tttaaaattt ttggcggctc tcaggctgct tacattttaa ccagttcagt
     5821 gaaaagttct ttttcagcaa atttctgttt agcaccatag ctaaaacttg cgtggaacat
     5881 attaagaatt gaccgaaatg acacaatctc aattatattt tttttgaaaa gttttcttta
     5941 tcaaatattt taaatcattg atttatatat aagtatacat tcattttaat aattaatctt
     6001 tatttaacaa tgatttatct atattcaatt gtttaattat tcttactaat attatctcta
     6061 tatcaatatt ttttatttaa aaacatatgt ttagtagtgc ttttgattaa agtaccagag
     6121 ggagggagca gagctgaatg ggaaatactc accctagagc gattcttaaa aatcacccta
     6181 aagtattccc attcgatgta ccgtcggtcg gtcgctttcg catcagggat gacatcactg
     6241 tatcaagctg ccactgttat gattacgatt gatagcaccg cctgaacacg ctcataaccg
     6301 ccaattaaat gactactcat tgcgtccgct actctgttca gttcccctag taatagcgtt
     6361 tttccgatgt gtgctagcgt cactgtacct catcacccac acatggacag attgagttac
     6421 agacattggc taaatttttg gggtctatgc ttgacaaagc gagttaaaac cttagaattt
     6481 aagacaggta cattaagccc ctgtggtgaa atcattagcg gtcgtcaaac cttaatgctt
     6541 tcgattgccg atatgtcagt atcgaagatc tgtaccccat aaattacggt aaagccccaa
     6601 gcaattgcaa ggggctttta tcttttttaa caaaaaaaat ttataaatca ggattttata
     6661 acactaaata atccaaagta catgagtaag tcatgaccac tcctgcgatg tgtgtagcac
     6721 tctgagtatc cgtattatat cagtgatgtc atatacaacc acataatgcg gatgtatcac
     6781 taactcccta gtatctttct gtctgcctgt gcgccccatc agtcgattat caacaagtaa
     6841 atctgtcttt tcttcaatta aatcatcaat ttcaatagct gctatagggt tgcgttcttc
     6901 aagataatca tagatatttc tacgatcttg gcgagctgtt tcagtccact caatcttcat
     6961 tcttcgctgc cagtttcgcc aatgttgcag cacgtctagc ggcaaactct gcctttacgt
     7021 cctcatgtgg aatcaacttg cctgcattgg cttcatttaa gccgatctgt acctgctgct
     7081 tgaaccattt gtcatagtct gcggcttcct gttgcttctt aacaaagtca cgcatgaagt
     7141 cacggattag ctgcgccccc gaccgatcaa cagtcttagc aaggcttgaa aattcttgct
     7201 ttaggtcatg gtctacccga aaggtaaaag tcgcttctgt catgtttttc actccaatgt
     7261 gttacatttg aatgctattg taacacatta aaaagttagg tcattgtctt ttttcggctt
     7321 aggctgttct tggtcataac gtggtttaaa tgatggctgt cgctcattct caagtaaaat
     7381 tttccgttta ttaacagcct gttgcatctg ctcatagtaa ggcatcaact gcggtaaaat
     7441 atcagcaagt gcctgtctgt ctttatccag gatacggctc aattgcccga tttttttcaa
     7501 gtcacgctca taggctattc tgtcgctgat ctgctcgtat ttttgctttg aacaccttat
     7561 gtactcttgc atatcgttac ttgcccatgc ctgcgcctga agacgtgtaa acatggaaat
     7621 gtctgaagca acatggtcgc tgatatggtc atactcagca tctttgacat agccgtattt
     7681 tgataaatat ctttcatagt cctgccgttt ctttttgtca tcttcctgtt tttgtttagt
     7741 ttctctttct atctgctgtt gttttcgttg ctgttcctgt tgctcctctt ttatctctct
     7801 gtgataaatg gaaaggctag attgatcatt atcccaacgt gtgagcatgt cctcaaagcc
     7861 gtaataatca gcactaaacc aacctgtttt cactttcact ggtggcggca gtggctctcc
     7921 gatctccctt agcagtgctg tagctgctcg tccctctttg atcactttat caagctgcaa
     7981 gcggtcgata ttgtcttgca actgcaggct tcctcgctca ctgactcgct gcgctcggtc
     8041 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     8101 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     8161 aaaaaggccg cgttgctggc gtttttccac aggctccgcc cccctgacga gcatcacaaa
     8221 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     8281 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     8341 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     8401 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     8461 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     8521 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     8581 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     8641 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     8701 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     8761 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     8821 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     8881 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     8941 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     9001 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     9061 cccagtgctg caatgatacc gcgagagcca cgctcaccgg ctccagattt atcagcaata
     9121 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     9181 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     9241 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     9301 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     9361 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     9421 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     9481 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     9541 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     9601 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     9661 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     9721 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     9781 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     9841 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     9901 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     9961 acattaacct ataaaaatag gcgtatcacg aggcagaatt tcagataaaa aaaatcctta
    10021 gctttcgcta aggatgattt ctggaattcg cggccgcttc tagaaagtat aggaacttca
    10081 gagcgctttt gaagctgatg tgcttaaaaa cttactcaat ggaattagct cctcgagcag
    10141 aaagtcaaaa gcctccgacc ggaggctttt gacttgaggg ggatcgatcc cttatggctc
    10201 tgcacccggc tccatcacca acaggtcgcg cacgcgcttc actcggttgc ggatcgacac
//
