LOCUS       sequence-404860-       10260 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 3 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: pMB3-B-TelR
FEATURES             Location/Qualifiers
     source          1..10260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    88..109
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(124..129,130..147,148..154)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    162..178
                     /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             196..207
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             214..891
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      936..1007
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1023..1050
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(5016..5033)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5187..5775)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5267..5286)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5946..6737,6738..6806))
                     /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     6569..6588
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6807..6911)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6979..6997
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(7001..7044)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    7047..7066
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     terminator      7146..7177
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     RBS             10078..10100
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
       61 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
      121 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
      181 tcacacatac tagagaaaga ggagaaatac tagatggctt cctccgaaga cgttatcaaa
      241 gagttcatgc gtttcaaagt tcgtatggaa ggttccgtta acggtcacga gttcgaaatc
      301 gaaggtgaag gtgaaggtcg tccgtacgaa ggtacccaga ccgctaaact gaaagttacc
      361 aaaggtggtc cgctgccgtt cgcttgggac atcctgtccc cgcagttcca gtacggttcc
      421 aaagcttacg ttaaacaccc ggctgacatc ccggactacc tgaaactgtc cttcccggaa
      481 ggtttcaaat gggaacgtgt tatgaacttc gaagacggtg gtgttgttac cgttacccag
      541 gactcctccc tgcaagacgg tgagttcatc tacaaagtta aactgcgtgg taccaacttc
      601 ccgtccgacg gtccggttat gcagaaaaaa accatgggtt gggaagcttc caccgaacgt
      661 atgtacccgg aagacggtgc tctgaaaggt gaaatcaaaa tgcgtctgaa actgaaagac
      721 ggtggtcact acgacgctga agttaaaacc acctacatgg ctaaaaaacc ggttcagctg
      781 ccgggtgctt acaaaaccga catcaaactg gacatcacct cccacaacga agactacacc
      841 atcgttgaac agtacgaacg tgctgaaggt cgtcactcca ccggtgctta ataacgctga
      901 tagtgctagt gtagatcgct actagagcca ggcatcaaat aaaacgaaag gctcagtcga
      961 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca
     1021 ctggctcacc ttcgggtggg cctttctgcg tttataggtc tcatgtttta gagagacgct
     1081 gcagttaaca agttgcctga cgctgtgtct tatccagttc atgcggcaat gctatctcta
     1141 cttcctttgc taatcttgcc tcctgtttga tgtcaccgtt cttttttagt tcggattgct
     1201 ctacccgatt ccatagggtt tgacgatcta acatgtcagg acttgcccca gttggggcaa
     1261 aaatttgggt gtattcaatg cctgtttttt tggtgtagtc ctgctctttt ccgtacgtat
     1321 cacagtacaa tttttcgcct gctcggtatg ctgcacatgc cacgattgag cgaccatctg
     1381 acctcgaaat gttctgcatt tcacaatggt aaattgccat tttcatcacc tgtttattaa
     1441 cagccgcccc aagttttggg cggttggggt gtcggggttt ccctgaccgt acttgcaaaa
     1501 tttggcgtag caaaaatttt gcgtaagtgc gcccttcggg aactctgcga ggctaaaagc
     1561 aaaagcaaga gcaataaggc tttgactttg ttttttgatt ttgctcgctg cgctcggatc
     1621 ttgatgtaag tcgagttttt gaagtagaac acttttcaca ttgatgatgg ttcatacttc
     1681 ggaaaatagg ttgagacaca ggcattaaaa atggtcaaaa aaacagctat ggaattgggt
     1741 caaatgctcg atgaagaaaa ggaaaaattg gaacgtcagg aaaaggcaag aaaggatcta
     1801 gctgacgctg tggtgaaagg tcgagagcaa aaacaggcaa ggtcagacga tgcaaaacgc
     1861 aagatcctga tcggggctta cttactgaag aaacatgata atgacatcaa gaaacttgtg
     1921 gaacaaaacc ccgattttat tgggtacatt agagaaaatg ataagcactt attttcatag
     1981 gcattagtag ttataaaaaa ccctcataaa tcgagggttt cttttttatc tgcattactt
     2041 tttgaatcaa tgtacttttt tagcttgcta tttttcactg catacagccg cttatactct
     2101 ttctcaatca atgcggtcaa aatggctttt tggctctccc ctgctgcgcc caccatgtcc
     2161 gcaagcatat ccgacacacc cttatctata aacacctcca aacgttgact atcaaggctt
     2221 ttgcgttttt cacgatatgc tttttgtcgt tcggcattgg tttttggcgg ctcttttagc
     2281 atgtcctgtg ttttctgatc gttcgggtct ttcatgtctt taactcccat atagtatatc
     2341 atagttacag gataacattg tttttagtta caggataact ttttataaaa tttatttgta
     2401 aaaatttatg ctatcctgca actattttta aattacagga taactattta acgttaccct
     2461 gtaactatag ttatcctgta acgctaaatt aattaccctg taacatatat tttgttatcc
     2521 tgtaacaact aaatattgaa ataatactac caaagttacc ctgtaacgaa aataactagg
     2581 ttaaaaaatg atcggatttt aacattttgc gttgttccaa aagttatcaa cagcctagaa
     2641 cgtcatagga agcgattaca gacactttat agctatcagc atgggaacat aagggcagga
     2701 tgaaatatgg gtcttaaaac gcaaatggtg aggttttaga ggtatttttt gaagatgatt
     2761 aaggcggttt gtttttaaaa tttttggcgg ctctcaggct gcttacattt taaccagttc
     2821 agtgaaaagt tctttttcag caaatttctg tttagcacca tagctaaaac ttgcgtggaa
     2881 catattaaga attgaccgaa atgacacaat ctcaattata ttttttttga aaagttttct
     2941 ttatcaaata ttttaaatca ttgatttata tataagtata cattcatttt aataattaat
     3001 ctttatttaa caatgattta tctatattca attgtttaat tattcttact aatattatct
     3061 ctatatcaat attttttatt taaaaacata tgtttagtag tgcttttgat taaagtacca
     3121 gagggaggga gcagagctga atgggaaata ctcaccctag agcgattctt aaaaatcacc
     3181 ctaaagtatt cccattcgat gtaccgtcgg tcggtcgctt tcgcatcagg gatgacatca
     3241 ctgtatcaag ctgccactgt tatgattacg attgatagca ccgcctgaac acgctcataa
     3301 ccgccaatta aatgactact cattgcgtcc gctactctgt tcagttcccc tagtaatagc
     3361 gtttttccga tgtgtgctag cgtcactgta cctcatcacc cacacatgga cagattgagt
     3421 tacagacatt ggctaaattt ttggggtcta tgcttgacaa agcgagttaa aaccttagaa
     3481 tttaagacag gtacattaag cccctgtggt gaaatcatta gcggtcgtca aaccttaatg
     3541 ctttcgattg ccgatatgtc agtatcgaag atctgtaccc cataaattac ggtaaagccc
     3601 caagcaattg caaggggctt ttatcttttt taacaaaaaa aatttataaa tcaggatttt
     3661 ataacactaa ataatccaaa gtacatgagt aagtcatgac cactcctgcg atgtgtgtag
     3721 cactctgagt atccgtatta tatcagtgat gtcatataca accacataat gcggatgtat
     3781 cactaactcc ctagtatctt tctgtctgcc tgtgcgcccc atcagtcgat tatcaacaag
     3841 taaatctgtc ttttcttcaa ttaaatcatc aatttcaata gctgctatag ggttgcgttc
     3901 ttcaagataa tcatagatat ttctacgatc ttggcgagct gtttcagtcc actcaatctt
     3961 cattcttcgc tgccagtttc gccaatgttg cagcacgtct agcggcaaac tctgccttta
     4021 cgtcctcatg tggaatcaac ttgcctgcat tggcttcatt taagccgatc tgtacctgct
     4081 gcttgaacca tttgtcatag tctgcggctt cctgttgctt cttaacaaag tcacgcatga
     4141 agtcacggat tagctgcgcc cccgaccgat caacagtctt agcaaggctt gaaaattctt
     4201 gctttaggtc atggtctacc cgaaaggtaa aagtcgcttc tgtcatgttt ttcactccaa
     4261 tgtgttacat ttgaatgcta ttgtaacaca ttaaaaagtt aggtcattgt cttttttcgg
     4321 cttaggctgt tcttggtcat aacgtggttt aaatgatggc tgtcgctcat tctcaagtaa
     4381 aattttccgt ttattaacag cctgttgcat ctgctcatag taaggcatca actgcggtaa
     4441 aatatcagca agtgcctgtc tgtctttatc caggatacgg ctcaattgcc cgattttttt
     4501 caagtcacgc tcataggcta ttctgtcgct gatctgctcg tatttttgct ttgaacacct
     4561 tatgtactct tgcatatcgt tacttgccca tgcctgcgcc tgaagacgtg taaacatgga
     4621 aatgtctgaa gcaacatggt cgctgatatg gtcatactca gcatctttga catagccgta
     4681 ttttgataaa tatctttcat agtcctgccg tttctttttg tcatcttcct gtttttgttt
     4741 agtttctctt tctatctgct gttgttttcg ttgctgttcc tgttgctcct cttttatctc
     4801 tctgtgataa atggaaaggc tagattgatc attatcccaa cgtgtgagca tgtcctcaaa
     4861 gccgtaataa tcagcactaa accaacctgt tttcactttc actggtggcg gcagtggctc
     4921 tccgatctcc cttagcagtg ctgtagctgc tcgtccctct ttgatcactt tatcaagctg
     4981 caagcggtcg atattgtctt gcaactgcag gcttcctcgc tcactgactc gctgcgctcg
     5041 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5101 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5161 cgtaaaaagg ccgcgttgct ggcgtttttc cacaggctcc gcccccctga cgagcatcac
     5221 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5281 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5341 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5401 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5461 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5521 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5581 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5641 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5701 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5761 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5821 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5881 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5941 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6001 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6061 ggccccagtg ctgcaatgat accgcgagag ccacgctcac cggctccaga tttatcagca
     6121 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6181 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6241 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6301 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6361 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6421 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6481 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6541 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6601 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6661 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6721 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6781 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6841 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6901 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     6961 atgacattaa cctataaaaa taggcgtatc acgaggcaga atttcagata aaaaaaatcc
     7021 ttagctttcg ctaaggatga tttctggaat tcgcggccgc ttctagaaag tataggaact
     7081 tcagagcgct tttgaagctg atgtgcttaa aaacttactc aatggaatta gctcctcgag
     7141 cagaaagtca aaagcctccg accggaggct tttgacttga gggggatcga tcccttatgg
     7201 ctctgcaccc ggctccatca ccaacaggtc gcgcacgcgc ttcactcggt tgcggatcga
     7261 cactgccagc ccaacaaagc cggttgccgc cgccgccagg atcgcgccga tgatgccggc
     7321 cacaccggcc atcgcccacc aggtcgccgc cttccggttc cattcctgct ggtactgctt
     7381 cgcaatgctg gacctcggct caccataggc tgaccgctcg atggcgtatg ccgcttctcc
     7441 ccttggcgta aaacccagcg ccgcaggcgg cattgccatg ctgcccgccg ctttcccgac
     7501 cacgacgcgc gcaccaggct tgcggtccag accttcggcc acggcgagct gcgcaaggac
     7561 ataatcagcc gccgacttgg ctccacgcgc ctcgatcagc tcttgcactc gcgcgaaatc
     7621 cttggcctcc acggccgcca tgaatcgcgc acgcggcgaa ggctccgcag ggccggcgtc
     7681 gtgatcgccg ccgagaatgc ccttcaccaa gttcgacgac acgaaaatca tgctgacggc
     7741 tatcaccatc atgcagacgg atcgcacgaa cccgcagaac tcacccccga acacgagcac
     7801 ggcacccgcg accactatgc caagaatgcc caaggtaaaa attgccggcc ccgccatgaa
     7861 gtccgtgaat gccccgacgg ccgaagtgaa gggcaggccg ccacccaggc cgccgccctc
     7921 actgcccggc acctggtcgc tgaatgtcga tgccagcacc tgcggcacgt caatgcttcc
     7981 gggcgtcgcg ctcgggctga tcgcccatcc cgttactgcc ccgatcccgg caatggcaag
     8041 gactgccagc gccgcgatga ggaagcgggt gccccgcttc ttcatcttcg cgcctcgggc
     8101 ctccaggccg cctacctggg cgaaaacatc ggtgtttgtg gcattcatac ggactcctgt
     8161 tgggccagct cgcgcacggg ctggcgggtc agcttggctt gaagatcgcc acgcattgcg
     8221 gcgatctgct tctcggcatc cttgcgcttc tgcacgcctt cctgctggat gcgaataacg
     8281 tcctcgacgg tcttgatgag cgtcgtctga acctgcttga gcgtgtccac gtcgatcacc
     8341 aggcgttggt tctccttcgc cgtttcgacg gacgtgcgat gcagcagggc cgcattgcgc
     8401 ttcatcaggt cgttggtggt gtcgtcgatg gccgtggcca gttcgacggc gttcttctgc
     8461 tcgttgaggc tcaaggccag catgaattgc cgcttccacg ccggcacggt gatttcgcgg
     8521 atggtgtgga atttatcgac cagcatctgg ttgttggcct ggatcatgcg gatggtcggc
     8581 aggctctgca tggccgaatg ttgcaaggcg atcaggtcgc cgatgcgctt gtccaggttg
     8641 gcaaccatcg catcgaggtc ggccagctcc tgcacgcggc cagggtcgtt cccgacattg
     8701 ccgcgcagac cctcggcctg ctcgcgcagc tcggcaaggc ggaccttgcc ggccgcgatg
     8761 tggacgccaa gaaggcggtg ttcctcgcgc acggctgcga acatttcgtc gagcgaggca
     8821 ttgcgctgcg cgatgccttg ctgggtggtc tgcacttcgc tgaccaggtg ttcgatctgc
     8881 tcgcgggtcg tgtcgaagcg cgccatgaag cccgtcgaac ggacgcggga gcggtcgatc
     8941 agcgggccaa tcaggggcag gcgggaacgg ttgtcggaca aagggccgac gttcagggaa
     9001 cgggccttgg cgacaacctg ggtcagtttc tcgcctgctt cgtccaggtc gctgttgcgc
     9061 acctggtcca gcaggctatc ggcgtagcgg gacgtgtgct cggccacgtc gcggccgaac
     9121 tcggcaacgg tctgcggact gccgacctcg atccgctgcg cgaccgcatg gacttccggc
     9181 acgtcgcttt cctgcaagcc cagctcgcgc agggttgccg gggtcatgtc aaaggcgacg
     9241 ataggggcct tggcgtcgtg cgtcgttttc agtgcgttca tagggttctc ccgccgtgtt
     9301 attggttgat gccttccagg ctctgcgaaa ggctccgcat gagcgcctgg tgagctttgg
     9361 ccgcctcggc gaccattgcc ggattcatgt tcttggtggt gatgagcgcg agggtgtgct
     9421 gacgccagac gggcaccagg acggatgccg tttcagagaa gcggtccagc atgtccacgg
     9481 cctgcgcccg cgtgagcttc atctgagtga cgctcatttc atgggacgcc atgagggttg
     9541 ccaggttggc gagcttgcgc gcgaagcgtt cgcgcggctt gtcgaactcg atcacgccgg
     9601 ccttggccgc gccggcctcg gggttctcgt ccaggaactc gcgcccggct tgaatgtagg
     9661 ctctgagccg gtctacctcg gcctcatgcg tattgagcat gtcatccaag gcgcgcaacg
     9721 tgtcccgcac gcgctgcgct acgccctcgg cttcgtccag caactggtcg agcgtcttgc
     9781 gggcgacctg atacctcacc tggcgttcaa cctcacggcc aagcatcttc tcgaaccagg
     9841 taggcttttc cgcgatcttg cgggggtccg cgtcgaccag cttcgccacg atctggctga
     9901 ttttgtcggc cagcgcggca actgcgccgt gctccatcag attcgacagc tcgttgaggg
     9961 aatccgcccc gtcgatgccg gccccgtact cgccaatcgt cgccggcgac gcgaagaggg
    10021 cgggcaaaac ctcccccttc aatcgcgcca tgttcacgct tgttcttcca tggtatatct
    10081 ccttcttaaa gttaaacaaa attattcgga acccagcatg atattccgga aataccaact
    10141 aagtcaacgg ctgatatcca ttgctgttga caaagggaat caggggatct tgaagttcct
    10201 attccgaagt tcctattctc tagaacgtct caatctctat agagacccaa tacgcaaacc
//
