LOCUS       sequence-404865-       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-D1-TelR
FEATURES             Location/Qualifiers
     source          1..10260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    212..233
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(248..253,254..271,272..278)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    286..302
                     /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             320..331
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             338..1015
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1060..1131
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1147..1174
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(5140..5157)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5311..5899)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5391..5410)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6070..6861,6862..6930))
                     /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     6693..6712
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6931..7035)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7103..7121
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(7125..7168)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    7171..7190
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     terminator      7270..7301
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     RBS             10202..10224
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 aactaagtca acggctgata tccattgctg ttgacaaagg gaatcagggg atcttgaagt
       61 tcctattccg aagttcctat tctctagaac gtctcagaag ctatagagac ccaatacgca
      121 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
      181 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
      241 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
      301 aatttcacac atactagaga aagaggagaa atactagatg gcttcctccg aagacgttat
      361 caaagagttc atgcgtttca aagttcgtat ggaaggttcc gttaacggtc acgagttcga
      421 aatcgaaggt gaaggtgaag gtcgtccgta cgaaggtacc cagaccgcta aactgaaagt
      481 taccaaaggt ggtccgctgc cgttcgcttg ggacatcctg tccccgcagt tccagtacgg
      541 ttccaaagct tacgttaaac acccggctga catcccggac tacctgaaac tgtccttccc
      601 ggaaggtttc aaatgggaac gtgttatgaa cttcgaagac ggtggtgttg ttaccgttac
      661 ccaggactcc tccctgcaag acggtgagtt catctacaaa gttaaactgc gtggtaccaa
      721 cttcccgtcc gacggtccgg ttatgcagaa aaaaaccatg ggttgggaag cttccaccga
      781 acgtatgtac ccggaagacg gtgctctgaa aggtgaaatc aaaatgcgtc tgaaactgaa
      841 agacggtggt cactacgacg ctgaagttaa aaccacctac atggctaaaa aaccggttca
      901 gctgccgggt gcttacaaaa ccgacatcaa actggacatc acctcccaca acgaagacta
      961 caccatcgtt gaacagtacg aacgtgctga aggtcgtcac tccaccggtg cttaataacg
     1021 ctgatagtgc tagtgtagat cgctactaga gccaggcatc aaataaaacg aaaggctcag
     1081 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt
     1141 cacactggct caccttcggg tgggcctttc tgcgtttata ggtctcatgt tcttcagaga
     1201 cgctgcagtt aacaagttgc ctgacgctgt gtcttatcca gttcatgcgg caatgctatc
     1261 tctacttcct ttgctaatct tgcctcctgt ttgatgtcac cgttcttttt tagttcggat
     1321 tgctctaccc gattccatag ggtttgacga tctaacatgt caggacttgc cccagttggg
     1381 gcaaaaattt gggtgtattc aatgcctgtt tttttggtgt agtcctgctc ttttccgtac
     1441 gtatcacagt acaatttttc gcctgctcgg tatgctgcac atgccacgat tgagcgacca
     1501 tctgacctcg aaatgttctg catttcacaa tggtaaattg ccattttcat cacctgttta
     1561 ttaacagccg ccccaagttt tgggcggttg gggtgtcggg gtttccctga ccgtacttgc
     1621 aaaatttggc gtagcaaaaa ttttgcgtaa gtgcgccctt cgggaactct gcgaggctaa
     1681 aagcaaaagc aagagcaata aggctttgac tttgtttttt gattttgctc gctgcgctcg
     1741 gatcttgatg taagtcgagt ttttgaagta gaacactttt cacattgatg atggttcata
     1801 cttcggaaaa taggttgaga cacaggcatt aaaaatggtc aaaaaaacag ctatggaatt
     1861 gggtcaaatg ctcgatgaag aaaaggaaaa attggaacgt caggaaaagg caagaaagga
     1921 tctagctgac gctgtggtga aaggtcgaga gcaaaaacag gcaaggtcag acgatgcaaa
     1981 acgcaagatc ctgatcgggg cttacttact gaagaaacat gataatgaca tcaagaaact
     2041 tgtggaacaa aaccccgatt ttattgggta cattagagaa aatgataagc acttattttc
     2101 ataggcatta gtagttataa aaaaccctca taaatcgagg gtttcttttt tatctgcatt
     2161 actttttgaa tcaatgtact tttttagctt gctatttttc actgcataca gccgcttata
     2221 ctctttctca atcaatgcgg tcaaaatggc tttttggctc tcccctgctg cgcccaccat
     2281 gtccgcaagc atatccgaca cacccttatc tataaacacc tccaaacgtt gactatcaag
     2341 gcttttgcgt ttttcacgat atgctttttg tcgttcggca ttggtttttg gcggctcttt
     2401 tagcatgtcc tgtgttttct gatcgttcgg gtctttcatg tctttaactc ccatatagta
     2461 tatcatagtt acaggataac attgttttta gttacaggat aactttttat aaaatttatt
     2521 tgtaaaaatt tatgctatcc tgcaactatt tttaaattac aggataacta tttaacgtta
     2581 ccctgtaact atagttatcc tgtaacgcta aattaattac cctgtaacat atattttgtt
     2641 atcctgtaac aactaaatat tgaaataata ctaccaaagt taccctgtaa cgaaaataac
     2701 taggttaaaa aatgatcgga ttttaacatt ttgcgttgtt ccaaaagtta tcaacagcct
     2761 agaacgtcat aggaagcgat tacagacact ttatagctat cagcatggga acataagggc
     2821 aggatgaaat atgggtctta aaacgcaaat ggtgaggttt tagaggtatt ttttgaagat
     2881 gattaaggcg gtttgttttt aaaatttttg gcggctctca ggctgcttac attttaacca
     2941 gttcagtgaa aagttctttt tcagcaaatt tctgtttagc accatagcta aaacttgcgt
     3001 ggaacatatt aagaattgac cgaaatgaca caatctcaat tatatttttt ttgaaaagtt
     3061 ttctttatca aatattttaa atcattgatt tatatataag tatacattca ttttaataat
     3121 taatctttat ttaacaatga tttatctata ttcaattgtt taattattct tactaatatt
     3181 atctctatat caatattttt tatttaaaaa catatgttta gtagtgcttt tgattaaagt
     3241 accagaggga gggagcagag ctgaatggga aatactcacc ctagagcgat tcttaaaaat
     3301 caccctaaag tattcccatt cgatgtaccg tcggtcggtc gctttcgcat cagggatgac
     3361 atcactgtat caagctgcca ctgttatgat tacgattgat agcaccgcct gaacacgctc
     3421 ataaccgcca attaaatgac tactcattgc gtccgctact ctgttcagtt cccctagtaa
     3481 tagcgttttt ccgatgtgtg ctagcgtcac tgtacctcat cacccacaca tggacagatt
     3541 gagttacaga cattggctaa atttttgggg tctatgcttg acaaagcgag ttaaaacctt
     3601 agaatttaag acaggtacat taagcccctg tggtgaaatc attagcggtc gtcaaacctt
     3661 aatgctttcg attgccgata tgtcagtatc gaagatctgt accccataaa ttacggtaaa
     3721 gccccaagca attgcaaggg gcttttatct tttttaacaa aaaaaattta taaatcagga
     3781 ttttataaca ctaaataatc caaagtacat gagtaagtca tgaccactcc tgcgatgtgt
     3841 gtagcactct gagtatccgt attatatcag tgatgtcata tacaaccaca taatgcggat
     3901 gtatcactaa ctccctagta tctttctgtc tgcctgtgcg ccccatcagt cgattatcaa
     3961 caagtaaatc tgtcttttct tcaattaaat catcaatttc aatagctgct atagggttgc
     4021 gttcttcaag ataatcatag atatttctac gatcttggcg agctgtttca gtccactcaa
     4081 tcttcattct tcgctgccag tttcgccaat gttgcagcac gtctagcggc aaactctgcc
     4141 tttacgtcct catgtggaat caacttgcct gcattggctt catttaagcc gatctgtacc
     4201 tgctgcttga accatttgtc atagtctgcg gcttcctgtt gcttcttaac aaagtcacgc
     4261 atgaagtcac ggattagctg cgcccccgac cgatcaacag tcttagcaag gcttgaaaat
     4321 tcttgcttta ggtcatggtc tacccgaaag gtaaaagtcg cttctgtcat gtttttcact
     4381 ccaatgtgtt acatttgaat gctattgtaa cacattaaaa agttaggtca ttgtcttttt
     4441 tcggcttagg ctgttcttgg tcataacgtg gtttaaatga tggctgtcgc tcattctcaa
     4501 gtaaaatttt ccgtttatta acagcctgtt gcatctgctc atagtaaggc atcaactgcg
     4561 gtaaaatatc agcaagtgcc tgtctgtctt tatccaggat acggctcaat tgcccgattt
     4621 ttttcaagtc acgctcatag gctattctgt cgctgatctg ctcgtatttt tgctttgaac
     4681 accttatgta ctcttgcata tcgttacttg cccatgcctg cgcctgaaga cgtgtaaaca
     4741 tggaaatgtc tgaagcaaca tggtcgctga tatggtcata ctcagcatct ttgacatagc
     4801 cgtattttga taaatatctt tcatagtcct gccgtttctt tttgtcatct tcctgttttt
     4861 gtttagtttc tctttctatc tgctgttgtt ttcgttgctg ttcctgttgc tcctctttta
     4921 tctctctgtg ataaatggaa aggctagatt gatcattatc ccaacgtgtg agcatgtcct
     4981 caaagccgta ataatcagca ctaaaccaac ctgttttcac tttcactggt ggcggcagtg
     5041 gctctccgat ctcccttagc agtgctgtag ctgctcgtcc ctctttgatc actttatcaa
     5101 gctgcaagcg gtcgatattg tcttgcaact gcaggcttcc tcgctcactg actcgctgcg
     5161 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5221 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5281 gaaccgtaaa aaggccgcgt tgctggcgtt tttccacagg ctccgccccc ctgacgagca
     5341 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5401 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5461 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5521 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5581 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5641 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5701 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5761 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5821 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5881 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5941 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6001 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6061 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6121 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6181 atctggcccc agtgctgcaa tgataccgcg agagccacgc tcaccggctc cagatttatc
     6241 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6301 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6361 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6421 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6481 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6541 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6601 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6661 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6721 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6781 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6841 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6901 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6961 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7021 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     7081 tatcatgaca ttaacctata aaaataggcg tatcacgagg cagaatttca gataaaaaaa
     7141 atccttagct ttcgctaagg atgatttctg gaattcgcgg ccgcttctag aaagtatagg
     7201 aacttcagag cgcttttgaa gctgatgtgc ttaaaaactt actcaatgga attagctcct
     7261 cgagcagaaa gtcaaaagcc tccgaccgga ggcttttgac ttgaggggga tcgatccctt
     7321 atggctctgc acccggctcc atcaccaaca ggtcgcgcac gcgcttcact cggttgcgga
     7381 tcgacactgc cagcccaaca aagccggttg ccgccgccgc caggatcgcg ccgatgatgc
     7441 cggccacacc ggccatcgcc caccaggtcg ccgccttccg gttccattcc tgctggtact
     7501 gcttcgcaat gctggacctc ggctcaccat aggctgaccg ctcgatggcg tatgccgctt
     7561 ctccccttgg cgtaaaaccc agcgccgcag gcggcattgc catgctgccc gccgctttcc
     7621 cgaccacgac gcgcgcacca ggcttgcggt ccagaccttc ggccacggcg agctgcgcaa
     7681 ggacataatc agccgccgac ttggctccac gcgcctcgat cagctcttgc actcgcgcga
     7741 aatccttggc ctccacggcc gccatgaatc gcgcacgcgg cgaaggctcc gcagggccgg
     7801 cgtcgtgatc gccgccgaga atgcccttca ccaagttcga cgacacgaaa atcatgctga
     7861 cggctatcac catcatgcag acggatcgca cgaacccgca gaactcaccc ccgaacacga
     7921 gcacggcacc cgcgaccact atgccaagaa tgcccaaggt aaaaattgcc ggccccgcca
     7981 tgaagtccgt gaatgccccg acggccgaag tgaagggcag gccgccaccc aggccgccgc
     8041 cctcactgcc cggcacctgg tcgctgaatg tcgatgccag cacctgcggc acgtcaatgc
     8101 ttccgggcgt cgcgctcggg ctgatcgccc atcccgttac tgccccgatc ccggcaatgg
     8161 caaggactgc cagcgccgcg atgaggaagc gggtgccccg cttcttcatc ttcgcgcctc
     8221 gggcctccag gccgcctacc tgggcgaaaa catcggtgtt tgtggcattc atacggactc
     8281 ctgttgggcc agctcgcgca cgggctggcg ggtcagcttg gcttgaagat cgccacgcat
     8341 tgcggcgatc tgcttctcgg catccttgcg cttctgcacg ccttcctgct ggatgcgaat
     8401 aacgtcctcg acggtcttga tgagcgtcgt ctgaacctgc ttgagcgtgt ccacgtcgat
     8461 caccaggcgt tggttctcct tcgccgtttc gacggacgtg cgatgcagca gggccgcatt
     8521 gcgcttcatc aggtcgttgg tggtgtcgtc gatggccgtg gccagttcga cggcgttctt
     8581 ctgctcgttg aggctcaagg ccagcatgaa ttgccgcttc cacgccggca cggtgatttc
     8641 gcggatggtg tggaatttat cgaccagcat ctggttgttg gcctggatca tgcggatggt
     8701 cggcaggctc tgcatggccg aatgttgcaa ggcgatcagg tcgccgatgc gcttgtccag
     8761 gttggcaacc atcgcatcga ggtcggccag ctcctgcacg cggccagggt cgttcccgac
     8821 attgccgcgc agaccctcgg cctgctcgcg cagctcggca aggcggacct tgccggccgc
     8881 gatgtggacg ccaagaaggc ggtgttcctc gcgcacggct gcgaacattt cgtcgagcga
     8941 ggcattgcgc tgcgcgatgc cttgctgggt ggtctgcact tcgctgacca ggtgttcgat
     9001 ctgctcgcgg gtcgtgtcga agcgcgccat gaagcccgtc gaacggacgc gggagcggtc
     9061 gatcagcggg ccaatcaggg gcaggcggga acggttgtcg gacaaagggc cgacgttcag
     9121 ggaacgggcc ttggcgacaa cctgggtcag tttctcgcct gcttcgtcca ggtcgctgtt
     9181 gcgcacctgg tccagcaggc tatcggcgta gcgggacgtg tgctcggcca cgtcgcggcc
     9241 gaactcggca acggtctgcg gactgccgac ctcgatccgc tgcgcgaccg catggacttc
     9301 cggcacgtcg ctttcctgca agcccagctc gcgcagggtt gccggggtca tgtcaaaggc
     9361 gacgataggg gccttggcgt cgtgcgtcgt tttcagtgcg ttcatagggt tctcccgccg
     9421 tgttattggt tgatgccttc caggctctgc gaaaggctcc gcatgagcgc ctggtgagct
     9481 ttggccgcct cggcgaccat tgccggattc atgttcttgg tggtgatgag cgcgagggtg
     9541 tgctgacgcc agacgggcac caggacggat gccgtttcag agaagcggtc cagcatgtcc
     9601 acggcctgcg cccgcgtgag cttcatctga gtgacgctca tttcatggga cgccatgagg
     9661 gttgccaggt tggcgagctt gcgcgcgaag cgttcgcgcg gcttgtcgaa ctcgatcacg
     9721 ccggccttgg ccgcgccggc ctcggggttc tcgtccagga actcgcgccc ggcttgaatg
     9781 taggctctga gccggtctac ctcggcctca tgcgtattga gcatgtcatc caaggcgcgc
     9841 aacgtgtccc gcacgcgctg cgctacgccc tcggcttcgt ccagcaactg gtcgagcgtc
     9901 ttgcgggcga cctgatacct cacctggcgt tcaacctcac ggccaagcat cttctcgaac
     9961 caggtaggct tttccgcgat cttgcggggg tccgcgtcga ccagcttcgc cacgatctgg
    10021 ctgattttgt cggccagcgc ggcaactgcg ccgtgctcca tcagattcga cagctcgttg
    10081 agggaatccg ccccgtcgat gccggccccg tactcgccaa tcgtcgccgg cgacgcgaag
    10141 agggcgggca aaacctcccc cttcaatcgc gccatgttca cgcttgttct tccatggtat
    10201 atctccttct taaagttaaa caaaattatt cggaacccag catgatattc cggaaatacc
//
