LOCUS       sequence-404861-       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-B-TelR
FEATURES             Location/Qualifiers
     source          1..10260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(248..279)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     protein_bind    487..508
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(523..528,529..546,547..553)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    561..577
                     /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             595..606
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             613..1290
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1335..1406
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1422..1449
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(5415..5432)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5586..6174)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5666..5685)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6345..7136,7137..7205))
                     /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     6968..6987
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7206..7310)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7378..7396
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(7400..7443)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    7446..7467
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     RBS             7585..7607
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 ccatcgagcg gtcagcctat ggtgagccga ggtccagcat tgcgaagcag taccagcagg
       61 aatggaaccg gaaggcggcg acctggtggg cgatggccgg tgtggccggc atcatcggcg
      121 cgatcctggc ggcggcggca accggctttg ttgggctggc agtgtcgatc cgcaaccgag
      181 tgaagcgcgt gcgcgacctg ttggtgatgg agccgggtgc agagccataa gggatcgatc
      241 cccctcaagt caaaagcctc cggtcggagg cttttgactt tctgctcgag gagctaattc
      301 cattgagtaa gtttttaagc acatcagctt caaaagcgct ctgaagttcc tatactttct
      361 agaacgtctc atgccctata gagacccaat acgcaaaccg cctctccccg cgcgttggcc
      421 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
      481 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
      541 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacatact agagaaagag
      601 gagaaatact agatggcttc ctccgaagac gttatcaaag agttcatgcg tttcaaagtt
      661 cgtatggaag gttccgttaa cggtcacgag ttcgaaatcg aaggtgaagg tgaaggtcgt
      721 ccgtacgaag gtacccagac cgctaaactg aaagttacca aaggtggtcc gctgccgttc
      781 gcttgggaca tcctgtcccc gcagttccag tacggttcca aagcttacgt taaacacccg
      841 gctgacatcc cggactacct gaaactgtcc ttcccggaag gtttcaaatg ggaacgtgtt
      901 atgaacttcg aagacggtgg tgttgttacc gttacccagg actcctccct gcaagacggt
      961 gagttcatct acaaagttaa actgcgtggt accaacttcc cgtccgacgg tccggttatg
     1021 cagaaaaaaa ccatgggttg ggaagcttcc accgaacgta tgtacccgga agacggtgct
     1081 ctgaaaggtg aaatcaaaat gcgtctgaaa ctgaaagacg gtggtcacta cgacgctgaa
     1141 gttaaaacca cctacatggc taaaaaaccg gttcagctgc cgggtgctta caaaaccgac
     1201 atcaaactgg acatcacctc ccacaacgaa gactacacca tcgttgaaca gtacgaacgt
     1261 gctgaaggtc gtcactccac cggtgcttaa taacgctgat agtgctagtg tagatcgcta
     1321 ctagagccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     1381 tctgttgttt gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc
     1441 ctttctgcgt ttataggtct catgttccgg agagacgctg cagttaacaa gttgcctgac
     1501 gctgtgtctt atccagttca tgcggcaatg ctatctctac ttcctttgct aatcttgcct
     1561 cctgtttgat gtcaccgttc ttttttagtt cggattgctc tacccgattc catagggttt
     1621 gacgatctaa catgtcagga cttgccccag ttggggcaaa aatttgggtg tattcaatgc
     1681 ctgttttttt ggtgtagtcc tgctcttttc cgtacgtatc acagtacaat ttttcgcctg
     1741 ctcggtatgc tgcacatgcc acgattgagc gaccatctga cctcgaaatg ttctgcattt
     1801 cacaatggta aattgccatt ttcatcacct gtttattaac agccgcccca agttttgggc
     1861 ggttggggtg tcggggtttc cctgaccgta cttgcaaaat ttggcgtagc aaaaattttg
     1921 cgtaagtgcg cccttcggga actctgcgag gctaaaagca aaagcaagag caataaggct
     1981 ttgactttgt tttttgattt tgctcgctgc gctcggatct tgatgtaagt cgagtttttg
     2041 aagtagaaca cttttcacat tgatgatggt tcatacttcg gaaaataggt tgagacacag
     2101 gcattaaaaa tggtcaaaaa aacagctatg gaattgggtc aaatgctcga tgaagaaaag
     2161 gaaaaattgg aacgtcagga aaaggcaaga aaggatctag ctgacgctgt ggtgaaaggt
     2221 cgagagcaaa aacaggcaag gtcagacgat gcaaaacgca agatcctgat cggggcttac
     2281 ttactgaaga aacatgataa tgacatcaag aaacttgtgg aacaaaaccc cgattttatt
     2341 gggtacatta gagaaaatga taagcactta ttttcatagg cattagtagt tataaaaaac
     2401 cctcataaat cgagggtttc ttttttatct gcattacttt ttgaatcaat gtactttttt
     2461 agcttgctat ttttcactgc atacagccgc ttatactctt tctcaatcaa tgcggtcaaa
     2521 atggcttttt ggctctcccc tgctgcgccc accatgtccg caagcatatc cgacacaccc
     2581 ttatctataa acacctccaa acgttgacta tcaaggcttt tgcgtttttc acgatatgct
     2641 ttttgtcgtt cggcattggt ttttggcggc tcttttagca tgtcctgtgt tttctgatcg
     2701 ttcgggtctt tcatgtcttt aactcccata tagtatatca tagttacagg ataacattgt
     2761 ttttagttac aggataactt tttataaaat ttatttgtaa aaatttatgc tatcctgcaa
     2821 ctatttttaa attacaggat aactatttaa cgttaccctg taactatagt tatcctgtaa
     2881 cgctaaatta attaccctgt aacatatatt ttgttatcct gtaacaacta aatattgaaa
     2941 taatactacc aaagttaccc tgtaacgaaa ataactaggt taaaaaatga tcggatttta
     3001 acattttgcg ttgttccaaa agttatcaac agcctagaac gtcataggaa gcgattacag
     3061 acactttata gctatcagca tgggaacata agggcaggat gaaatatggg tcttaaaacg
     3121 caaatggtga ggttttagag gtattttttg aagatgatta aggcggtttg tttttaaaat
     3181 ttttggcggc tctcaggctg cttacatttt aaccagttca gtgaaaagtt ctttttcagc
     3241 aaatttctgt ttagcaccat agctaaaact tgcgtggaac atattaagaa ttgaccgaaa
     3301 tgacacaatc tcaattatat tttttttgaa aagttttctt tatcaaatat tttaaatcat
     3361 tgatttatat ataagtatac attcatttta ataattaatc tttatttaac aatgatttat
     3421 ctatattcaa ttgtttaatt attcttacta atattatctc tatatcaata ttttttattt
     3481 aaaaacatat gtttagtagt gcttttgatt aaagtaccag agggagggag cagagctgaa
     3541 tgggaaatac tcaccctaga gcgattctta aaaatcaccc taaagtattc ccattcgatg
     3601 taccgtcggt cggtcgcttt cgcatcaggg atgacatcac tgtatcaagc tgccactgtt
     3661 atgattacga ttgatagcac cgcctgaaca cgctcataac cgccaattaa atgactactc
     3721 attgcgtccg ctactctgtt cagttcccct agtaatagcg tttttccgat gtgtgctagc
     3781 gtcactgtac ctcatcaccc acacatggac agattgagtt acagacattg gctaaatttt
     3841 tggggtctat gcttgacaaa gcgagttaaa accttagaat ttaagacagg tacattaagc
     3901 ccctgtggtg aaatcattag cggtcgtcaa accttaatgc tttcgattgc cgatatgtca
     3961 gtatcgaaga tctgtacccc ataaattacg gtaaagcccc aagcaattgc aaggggcttt
     4021 tatctttttt aacaaaaaaa atttataaat caggatttta taacactaaa taatccaaag
     4081 tacatgagta agtcatgacc actcctgcga tgtgtgtagc actctgagta tccgtattat
     4141 atcagtgatg tcatatacaa ccacataatg cggatgtatc actaactccc tagtatcttt
     4201 ctgtctgcct gtgcgcccca tcagtcgatt atcaacaagt aaatctgtct tttcttcaat
     4261 taaatcatca atttcaatag ctgctatagg gttgcgttct tcaagataat catagatatt
     4321 tctacgatct tggcgagctg tttcagtcca ctcaatcttc attcttcgct gccagtttcg
     4381 ccaatgttgc agcacgtcta gcggcaaact ctgcctttac gtcctcatgt ggaatcaact
     4441 tgcctgcatt ggcttcattt aagccgatct gtacctgctg cttgaaccat ttgtcatagt
     4501 ctgcggcttc ctgttgcttc ttaacaaagt cacgcatgaa gtcacggatt agctgcgccc
     4561 ccgaccgatc aacagtctta gcaaggcttg aaaattcttg ctttaggtca tggtctaccc
     4621 gaaaggtaaa agtcgcttct gtcatgtttt tcactccaat gtgttacatt tgaatgctat
     4681 tgtaacacat taaaaagtta ggtcattgtc ttttttcggc ttaggctgtt cttggtcata
     4741 acgtggttta aatgatggct gtcgctcatt ctcaagtaaa attttccgtt tattaacagc
     4801 ctgttgcatc tgctcatagt aaggcatcaa ctgcggtaaa atatcagcaa gtgcctgtct
     4861 gtctttatcc aggatacggc tcaattgccc gatttttttc aagtcacgct cataggctat
     4921 tctgtcgctg atctgctcgt atttttgctt tgaacacctt atgtactctt gcatatcgtt
     4981 acttgcccat gcctgcgcct gaagacgtgt aaacatggaa atgtctgaag caacatggtc
     5041 gctgatatgg tcatactcag catctttgac atagccgtat tttgataaat atctttcata
     5101 gtcctgccgt ttctttttgt catcttcctg tttttgttta gtttctcttt ctatctgctg
     5161 ttgttttcgt tgctgttcct gttgctcctc ttttatctct ctgtgataaa tggaaaggct
     5221 agattgatca ttatcccaac gtgtgagcat gtcctcaaag ccgtaataat cagcactaaa
     5281 ccaacctgtt ttcactttca ctggtggcgg cagtggctct ccgatctccc ttagcagtgc
     5341 tgtagctgct cgtccctctt tgatcacttt atcaagctgc aagcggtcga tattgtcttg
     5401 caactgcagg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5461 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5521 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5581 gcgtttttcc acaggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5641 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5701 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5761 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5821 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5881 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5941 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6001 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     6061 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6121 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6181 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6241 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6301 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6361 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6421 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6481 ccgcgagagc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6541 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6601 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6661 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6721 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6781 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6841 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6901 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6961 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7021 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7081 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7141 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7201 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7261 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7321 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     7381 aggcgtatca cgaggcagaa tttcagataa aaaaaatcct tagctttcgc taaggatgat
     7441 ttctggaatt cgcggccgct tctagagaat aggaacttcg gaataggaac ttcaagatcc
     7501 cctgattccc tttgtcaaca gcaatggata tcagccgttg acttagttgg tatttccgga
     7561 atatcatgct gggttccgaa taattttgtt taactttaag aaggagatat accatggaag
     7621 aacaagcgtg aacatggcgc gattgaaggg ggaggttttg cccgccctct tcgcgtcgcc
     7681 ggcgacgatt ggcgagtacg gggccggcat cgacggggcg gattccctca acgagctgtc
     7741 gaatctgatg gagcacggcg cagttgccgc gctggccgac aaaatcagcc agatcgtggc
     7801 gaagctggtc gacgcggacc cccgcaagat cgcggaaaag cctacctggt tcgagaagat
     7861 gcttggccgt gaggttgaac gccaggtgag gtatcaggtc gcccgcaaga cgctcgacca
     7921 gttgctggac gaagccgagg gcgtagcgca gcgcgtgcgg gacacgttgc gcgccttgga
     7981 tgacatgctc aatacgcatg aggccgaggt agaccggctc agagcctaca ttcaagccgg
     8041 gcgcgagttc ctggacgaga accccgaggc cggcgcggcc aaggccggcg tgatcgagtt
     8101 cgacaagccg cgcgaacgct tcgcgcgcaa gctcgccaac ctggcaaccc tcatggcgtc
     8161 ccatgaaatg agcgtcactc agatgaagct cacgcgggcg caggccgtgg acatgctgga
     8221 ccgcttctct gaaacggcat ccgtcctggt gcccgtctgg cgtcagcaca ccctcgcgct
     8281 catcaccacc aagaacatga atccggcaat ggtcgccgag gcggccaaag ctcaccaggc
     8341 gctcatgcgg agcctttcgc agagcctgga aggcatcaac caataacacg gcgggagaac
     8401 cctatgaacg cactgaaaac gacgcacgac gccaaggccc ctatcgtcgc ctttgacatg
     8461 accccggcaa ccctgcgcga gctgggcttg caggaaagcg acgtgccgga agtccatgcg
     8521 gtcgcgcagc ggatcgaggt cggcagtccg cagaccgttg ccgagttcgg ccgcgacgtg
     8581 gccgagcaca cgtcccgcta cgccgatagc ctgctggacc aggtgcgcaa cagcgacctg
     8641 gacgaagcag gcgagaaact gacccaggtt gtcgccaagg cccgttccct gaacgtcggc
     8701 cctttgtccg acaaccgttc ccgcctgccc ctgattggcc cgctgatcga ccgctcccgc
     8761 gtccgttcga cgggcttcat ggcgcgcttc gacacgaccc gcgagcagat cgaacacctg
     8821 gtcagcgaag tgcagaccac ccagcaaggc atcgcgcagc gcaatgcctc gctcgacgaa
     8881 atgttcgcag ccgtgcgcga ggaacaccgc cttcttggcg tccacatcgc ggccggcaag
     8941 gtccgccttg ccgagctgcg cgagcaggcc gagggtctgc gcggcaatgt cgggaacgac
     9001 cctggccgcg tgcaggagct ggccgacctc gatgcgatgg ttgccaacct ggacaagcgc
     9061 atcggcgacc tgatcgcctt gcaacattcg gccatgcaga gcctgccgac catccgcatg
     9121 atccaggcca acaaccagat gctggtcgat aaattccaca ccatccgcga aatcaccgtg
     9181 ccggcgtgga agcggcaatt catgctggcc ttgagcctca acgagcagaa gaacgccgtc
     9241 gaactggcca cggccatcga cgacaccacc aacgacctga tgaagcgcaa tgcggccctg
     9301 ctgcatcgca cgtccgtcga aacggcgaag gagaaccaac gcctggtgat cgacgtggac
     9361 acgctcaagc aggttcagac gacgctcatc aagaccgtcg aggacgttat tcgcatccag
     9421 caggaaggcg tgcagaagcg caaggatgcc gagaagcaga tcgccgcaat gcgtggcgat
     9481 cttcaagcca agctgacccg ccagcccgtg cgcgagctgg cccaacagga gtccgtatga
     9541 atgccacaaa caccgatgtt ttcgcccagg taggcggcct ggaggcccga ggcgcgaaga
     9601 tgaagaagcg gggcacccgc ttcctcatcg cggcgctggc agtccttgcc attgccggga
     9661 tcggggcagt aacgggatgg gcgatcagcc cgagcgcgac gcccggaagc attgacgtgc
     9721 cgcaggtgct ggcatcgaca ttcagcgacc aggtgccggg cagtgagggc ggcggcctgg
     9781 gtggcggcct gcccttcact tcggccgtcg gggcattcac ggacttcatg gcggggccgg
     9841 caatttttac cttgggcatt cttggcatag tggtcgcggg tgccgtgctc gtgttcgggg
     9901 gtgagttctg cgggttcgtg cgatccgtct gcatgatggt gatagccgtc agcatgattt
     9961 tcgtgtcgtc gaacttggtg aagggcattc tcggcggcga tcacgacgcc ggccctgcgg
    10021 agccttcgcc gcgtgcgcga ttcatggcgg ccgtggaggc caaggatttc gcgcgagtgc
    10081 aagagctgat cgaggcgcgt ggagccaagt cggcggctga ttatgtcctt gcgcagctcg
    10141 ccgtggccga aggtctggac cgcaagcctg gtgcgcgcgt cgtggtcggg aaagcggcgg
    10201 gcagcatggc aatgccgcct gcggcgctgg gttttacgcc aaggggagaa gcggcatacg
//
