LOCUS       sequence-404863-       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-D-TelR
FEATURES             Location/Qualifiers
     source          1..10260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     224..243
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(462..566)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     634..652
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(656..699)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    702..721
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     terminator      801..832
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     RBS             3733..3755
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4003..4024
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4039..4044,4045..4062,4063..4069)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4077..4093
                     /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             4111..4122
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4129..4806
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      4851..4922
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4938..4965
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(8931..8948)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9102..9690)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9182..9201)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9861..10260,1..392,393..461))
                     /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"
ORIGIN
        1 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
       61 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
      121 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
      181 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
      241 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
      301 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
      361 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
      421 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
      481 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
      541 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
      601 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gcagaatttc
      661 agataaaaaa aatccttagc tttcgctaag gatgatttct ggaattcgcg gccgcttcta
      721 gaaagtatag gaacttcaga gcgcttttga agctgatgtg cttaaaaact tactcaatgg
      781 aattagctcc tcgagcagaa agtcaaaagc ctccgaccgg aggcttttga cttgaggggg
      841 atcgatccct tatggctctg cacccggctc catcaccaac aggtcgcgca cgcgcttcac
      901 tcggttgcgg atcgacactg ccagcccaac aaagccggtt gccgccgccg ccaggatcgc
      961 gccgatgatg ccggccacac cggccatcgc ccaccaggtc gccgccttcc ggttccattc
     1021 ctgctggtac tgcttcgcaa tgctggacct cggctcacca taggctgacc gctcgatggc
     1081 gtatgccgct tctccccttg gcgtaaaacc cagcgccgca ggcggcattg ccatgctgcc
     1141 cgccgctttc ccgaccacga cgcgcgcacc aggcttgcgg tccagacctt cggccacggc
     1201 gagctgcgca aggacataat cagccgccga cttggctcca cgcgcctcga tcagctcttg
     1261 cactcgcgcg aaatccttgg cctccacggc cgccatgaat cgcgcacgcg gcgaaggctc
     1321 cgcagggccg gcgtcgtgat cgccgccgag aatgcccttc accaagttcg acgacacgaa
     1381 aatcatgctg acggctatca ccatcatgca gacggatcgc acgaacccgc agaactcacc
     1441 cccgaacacg agcacggcac ccgcgaccac tatgccaaga atgcccaagg taaaaattgc
     1501 cggccccgcc atgaagtccg tgaatgcccc gacggccgaa gtgaagggca ggccgccacc
     1561 caggccgccg ccctcactgc ccggcacctg gtcgctgaat gtcgatgcca gcacctgcgg
     1621 cacgtcaatg cttccgggcg tcgcgctcgg gctgatcgcc catcccgtta ctgccccgat
     1681 cccggcaatg gcaaggactg ccagcgccgc gatgaggaag cgggtgcccc gcttcttcat
     1741 cttcgcgcct cgggcctcca ggccgcctac ctgggcgaaa acatcggtgt ttgtggcatt
     1801 catacggact cctgttgggc cagctcgcgc acgggctggc gggtcagctt ggcttgaaga
     1861 tcgccacgca ttgcggcgat ctgcttctcg gcatccttgc gcttctgcac gccttcctgc
     1921 tggatgcgaa taacgtcctc gacggtcttg atgagcgtcg tctgaacctg cttgagcgtg
     1981 tccacgtcga tcaccaggcg ttggttctcc ttcgccgttt cgacggacgt gcgatgcagc
     2041 agggccgcat tgcgcttcat caggtcgttg gtggtgtcgt cgatggccgt ggccagttcg
     2101 acggcgttct tctgctcgtt gaggctcaag gccagcatga attgccgctt ccacgccggc
     2161 acggtgattt cgcggatggt gtggaattta tcgaccagca tctggttgtt ggcctggatc
     2221 atgcggatgg tcggcaggct ctgcatggcc gaatgttgca aggcgatcag gtcgccgatg
     2281 cgcttgtcca ggttggcaac catcgcatcg aggtcggcca gctcctgcac gcggccaggg
     2341 tcgttcccga cattgccgcg cagaccctcg gcctgctcgc gcagctcggc aaggcggacc
     2401 ttgccggccg cgatgtggac gccaagaagg cggtgttcct cgcgcacggc tgcgaacatt
     2461 tcgtcgagcg aggcattgcg ctgcgcgatg ccttgctggg tggtctgcac ttcgctgacc
     2521 aggtgttcga tctgctcgcg ggtcgtgtcg aagcgcgcca tgaagcccgt cgaacggacg
     2581 cgggagcggt cgatcagcgg gccaatcagg ggcaggcggg aacggttgtc ggacaaaggg
     2641 ccgacgttca gggaacgggc cttggcgaca acctgggtca gtttctcgcc tgcttcgtcc
     2701 aggtcgctgt tgcgcacctg gtccagcagg ctatcggcgt agcgggacgt gtgctcggcc
     2761 acgtcgcggc cgaactcggc aacggtctgc ggactgccga cctcgatccg ctgcgcgacc
     2821 gcatggactt ccggcacgtc gctttcctgc aagcccagct cgcgcagggt tgccggggtc
     2881 atgtcaaagg cgacgatagg ggccttggcg tcgtgcgtcg ttttcagtgc gttcataggg
     2941 ttctcccgcc gtgttattgg ttgatgcctt ccaggctctg cgaaaggctc cgcatgagcg
     3001 cctggtgagc tttggccgcc tcggcgacca ttgccggatt catgttcttg gtggtgatga
     3061 gcgcgagggt gtgctgacgc cagacgggca ccaggacgga tgccgtttca gagaagcggt
     3121 ccagcatgtc cacggcctgc gcccgcgtga gcttcatctg agtgacgctc atttcatggg
     3181 acgccatgag ggttgccagg ttggcgagct tgcgcgcgaa gcgttcgcgc ggcttgtcga
     3241 actcgatcac gccggccttg gccgcgccgg cctcggggtt ctcgtccagg aactcgcgcc
     3301 cggcttgaat gtaggctctg agccggtcta cctcggcctc atgcgtattg agcatgtcat
     3361 ccaaggcgcg caacgtgtcc cgcacgcgct gcgctacgcc ctcggcttcg tccagcaact
     3421 ggtcgagcgt cttgcgggcg acctgatacc tcacctggcg ttcaacctca cggccaagca
     3481 tcttctcgaa ccaggtaggc ttttccgcga tcttgcgggg gtccgcgtcg accagcttcg
     3541 ccacgatctg gctgattttg tcggccagcg cggcaactgc gccgtgctcc atcagattcg
     3601 acagctcgtt gagggaatcc gccccgtcga tgccggcccc gtactcgcca atcgtcgccg
     3661 gcgacgcgaa gagggcgggc aaaacctccc ccttcaatcg cgccatgttc acgcttgttc
     3721 ttccatggta tatctccttc ttaaagttaa acaaaattat tcggaaccca gcatgatatt
     3781 ccggaaatac caactaagtc aacggctgat atccattgct gttgacaaag ggaatcaggg
     3841 gatcttgaag ttcctattcc gaagttccta ttctctagaa cgtctcagaa gctatagaga
     3901 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     3961 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     4021 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     4081 gagcggataa caatttcaca catactagag aaagaggaga aatactagat ggcttcctcc
     4141 gaagacgtta tcaaagagtt catgcgtttc aaagttcgta tggaaggttc cgttaacggt
     4201 cacgagttcg aaatcgaagg tgaaggtgaa ggtcgtccgt acgaaggtac ccagaccgct
     4261 aaactgaaag ttaccaaagg tggtccgctg ccgttcgctt gggacatcct gtccccgcag
     4321 ttccagtacg gttccaaagc ttacgttaaa cacccggctg acatcccgga ctacctgaaa
     4381 ctgtccttcc cggaaggttt caaatgggaa cgtgttatga acttcgaaga cggtggtgtt
     4441 gttaccgtta cccaggactc ctccctgcaa gacggtgagt tcatctacaa agttaaactg
     4501 cgtggtacca acttcccgtc cgacggtccg gttatgcaga aaaaaaccat gggttgggaa
     4561 gcttccaccg aacgtatgta cccggaagac ggtgctctga aaggtgaaat caaaatgcgt
     4621 ctgaaactga aagacggtgg tcactacgac gctgaagtta aaaccaccta catggctaaa
     4681 aaaccggttc agctgccggg tgcttacaaa accgacatca aactggacat cacctcccac
     4741 aacgaagact acaccatcgt tgaacagtac gaacgtgctg aaggtcgtca ctccaccggt
     4801 gcttaataac gctgatagtg ctagtgtaga tcgctactag agccaggcat caaataaaac
     4861 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     4921 tctactagag tcacactggc tcaccttcgg gtgggccttt ctgcgtttat aggtctcatg
     4981 ttttagagag acgctgcagt tgcaagacaa tatcgaccgc ttgcagcttg ataaagtgat
     5041 caaagaggga cgagcagcta cagcactgct aagggagatc ggagagccac tgccgccacc
     5101 agtgaaagtg aaaacaggtt ggtttagtgc tgattattac ggctttgagg acatgctcac
     5161 acgttgggat aatgatcaat ctagcctttc catttatcac agagagataa aagaggagca
     5221 acaggaacag caacgaaaac aacagcagat agaaagagaa actaaacaaa aacaggaaga
     5281 tgacaaaaag aaacggcagg actatgaaag atatttatca aaatacggct atgtcaaaga
     5341 tgctgagtat gaccatatca gcgaccatgt tgcttcagac atttccatgt ttacacgtct
     5401 tcaggcgcag gcatgggcaa gtaacgatat gcaagagtac ataaggtgtt caaagcaaaa
     5461 atacgagcag atcagcgaca gaatagccta tgagcgtgac ttgaaaaaaa tcgggcaatt
     5521 gagccgtatc ctggataaag acagacaggc acttgctgat attttaccgc agttgatgcc
     5581 ttactatgag cagatgcaac aggctgttaa taaacggaaa attttacttg agaatgagcg
     5641 acagccatca tttaaaccac gttatgacca agaacagcct aagccgaaaa aagacaatga
     5701 cctaactttt taatgtgtta caatagcatt caaatgtaac acattggagt gaaaaacatg
     5761 acagaagcga cttttacctt tcgggtagac catgacctaa agcaagaatt ttcaagcctt
     5821 gctaagactg ttgatcggtc gggggcgcag ctaatccgtg acttcatgcg tgactttgtt
     5881 aagaagcaac aggaagccgc agactatgac aaatggttca agcagcaggt acagatcggc
     5941 ttaaatgaag ccaatgcagg caagttgatt ccacatgagg acgtaaaggc agagtttgcc
     6001 gctagacgtg ctgcaacatt ggcgaaactg gcagcgaaga atgaagattg agtggactga
     6061 aacagctcgc caagatcgta gaaatatcta tgattatctt gaagaacgca accctatagc
     6121 agctattgaa attgatgatt taattgaaga aaagacagat ttacttgttg ataatcgact
     6181 gatggggcgc acaggcagac agaaagatac tagggagtta gtgatacatc cgcattatgt
     6241 ggttgtatat gacatcactg atataatacg gatactcaga gtgctacaca catcgcagga
     6301 gtggtcatga cttactcatg tactttggat tatttagtgt tataaaatcc tgatttataa
     6361 attttttttg ttaaaaaaga taaaagcccc ttgcaattgc ttggggcttt accgtaattt
     6421 atggggtaca gatcttcgat actgacatat cggcaatcga aagcattaag gtttgacgac
     6481 cgctaatgat ttcaccacag gggcttaatg tacctgtctt aaattctaag gttttaactc
     6541 gctttgtcaa gcatagaccc caaaaattta gccaatgtct gtaactcaat ctgtccatgt
     6601 gtgggtgatg aggtacagtg acgctagcac acatcggaaa aacgctatta ctaggggaac
     6661 tgaacagagt agcggacgca atgagtagtc atttaattgg cggttatgag cgtgttcagg
     6721 cggtgctatc aatcgtaatc ataacagtgg cagcttgata cagtgatgtc atccctgatg
     6781 cgaaagcgac cgaccgacgg tacatcgaat gggaatactt tagggtgatt tttaagaatc
     6841 gctctagggt gagtatttcc cattcagctc tgctccctcc ctctggtact ttaatcaaaa
     6901 gcactactaa acatatgttt ttaaataaaa aatattgata tagagataat attagtaaga
     6961 ataattaaac aattgaatat agataaatca ttgttaaata aagattaatt attaaaatga
     7021 atgtatactt atatataaat caatgattta aaatatttga taaagaaaac ttttcaaaaa
     7081 aaatataatt gagattgtgt catttcggtc aattcttaat atgttccacg caagttttag
     7141 ctatggtgct aaacagaaat ttgctgaaaa agaacttttc actgaactgg ttaaaatgta
     7201 agcagcctga gagccgccaa aaattttaaa aacaaaccgc cttaatcatc ttcaaaaaat
     7261 acctctaaaa cctcaccatt tgcgttttaa gacccatatt tcatcctgcc cttatgttcc
     7321 catgctgata gctataaagt gtctgtaatc gcttcctatg acgttctagg ctgttgataa
     7381 cttttggaac aacgcaaaat gttaaaatcc gatcattttt taacctagtt attttcgtta
     7441 cagggtaact ttggtagtat tatttcaata tttagttgtt acaggataac aaaatatatg
     7501 ttacagggta attaatttag cgttacagga taactatagt tacagggtaa cgttaaatag
     7561 ttatcctgta atttaaaaat agttgcagga tagcataaat ttttacaaat aaattttata
     7621 aaaagttatc ctgtaactaa aaacaatgtt atcctgtaac tatgatatac tatatgggag
     7681 ttaaagacat gaaagacccg aacgatcaga aaacacagga catgctaaaa gagccgccaa
     7741 aaaccaatgc cgaacgacaa aaagcatatc gtgaaaaacg caaaagcctt gatagtcaac
     7801 gtttggaggt gtttatagat aagggtgtgt cggatatgct tgcggacatg gtgggcgcag
     7861 caggggagag ccaaaaagcc attttgaccg cattgattga gaaagagtat aagcggctgt
     7921 atgcagtgaa aaatagcaag ctaaaaaagt acattgattc aaaaagtaat gcagataaaa
     7981 aagaaaccct cgatttatga gggtttttta taactactaa tgcctatgaa aataagtgct
     8041 tatcattttc tctaatgtac ccaataaaat cggggttttg ttccacaagt ttcttgatgt
     8101 cattatcatg tttcttcagt aagtaagccc cgatcaggat cttgcgtttt gcatcgtctg
     8161 accttgcctg tttttgctct cgacctttca ccacagcgtc agctagatcc tttcttgcct
     8221 tttcctgacg ttccaatttt tccttttctt catcgagcat ttgacccaat tccatagctg
     8281 tttttttgac catttttaat gcctgtgtct caacctattt tccgaagtat gaaccatcat
     8341 caatgtgaaa agtgttctac ttcaaaaact cgacttacat caagatccga gcgcagcgag
     8401 caaaatcaaa aaacaaagtc aaagccttat tgctcttgct tttgctttta gcctcgcaga
     8461 gttcccgaag ggcgcactta cgcaaaattt ttgctacgcc aaattttgca agtacggtca
     8521 gggaaacccc gacaccccaa ccgcccaaaa cttggggcgg ctgttaataa acaggtgatg
     8581 aaaatggcaa tttaccattg tgaaatgcag aacatttcga ggtcagatgg tcgctcaatc
     8641 gtggcatgtg cagcataccg agcaggcgaa aaattgtact gtgatacgta cggaaaagag
     8701 caggactaca ccaaaaaaac aggcattgaa tacacccaaa tttttgcccc aactggggca
     8761 agtcctgaca tgttagatcg tcaaacccta tggaatcggg tagagcaatc cgaactaaaa
     8821 aagaacggtg acatcaaaca ggaggcaaga ttagcaaagg aagtagagat agcattgccg
     8881 catgaactgg ataagacaca gcgtcaggca acttgttaac tgcaggcttc ctcgctcact
     8941 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     9001 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     9061 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccacag gctccgcccc
     9121 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     9181 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     9241 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     9301 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     9361 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     9421 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     9481 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     9541 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     9601 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     9661 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     9721 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     9781 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     9841 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     9901 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     9961 gagggcttac catctggccc cagtgctgca atgataccgc gagagccacg ctcaccggct
    10021 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
    10081 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
    10141 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
    10201 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
//
