LOCUS       sequence-404884-       10266 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 10266)
  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 10266)
  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-C-TelR
FEATURES             Location/Qualifiers
     source          1..10266
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(648..679)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     protein_bind    887..908
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(923..928,929..946,947..953)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    961..977
                     /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             995..1006
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1013..1690
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1735..1806
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1822..1849
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(5821..5838)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5992..6580)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6072..6091)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6751..7542,7543..7611))
                     /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     7374..7393
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7612..7716)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7784..7802
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(7806..7849)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    7852..7873
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     RBS             7991..8013
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 cttggcatag tggtcgcggg tgccgtgctc gtgttcgggg gtgagttctg cgggttcgtg
       61 cgatccgtct gcatgatggt gatagccgtc agcatgattt tcgtgtcgtc gaacttggtg
      121 aagggcattc tcggcggcga tcacgacgcc ggccctgcgg agccttcgcc gcgtgcgcga
      181 ttcatggcgg ccgtggaggc caaggatttc gcgcgagtgc aagagctgat cgaggcgcgt
      241 ggagccaagt cggcggctga ttatgtcctt gcgcagctcg ccgtggccga aggtctggac
      301 cgcaagcctg gtgcgcgcgt cgtggtcggg aaagcggcgg gcagcatggc aatgccgcct
      361 gcggcgctgg gttttacgcc aaggggagaa gcggcatacg ccatcgagcg gtcagcctat
      421 ggtgagccga ggtccagcat tgcgaagcag taccagcagg aatggaaccg gaaggcggcg
      481 acctggtggg cgatggccgg tgtggccggc atcatcggcg cgatcctggc ggcggcggca
      541 accggctttg ttgggctggc agtgtcgatc cgcaaccgag tgaagcgcgt gcgcgacctg
      601 ttggtgatgg agccgggtgc agagccataa gggatcgatc cccctcaagt caaaagcctc
      661 cggtcggagg cttttgactt tctgctcgag gagctaattc cattgagtaa gtttttaagc
      721 acatcagctt caaaagcgct ctgaagttcc tatactttct agaacgtctc accggctata
      781 gagacccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
      841 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      901 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      961 ttgtgagcgg ataacaattt cacacatact agagaaagag gagaaatact agatggcttc
     1021 ctccgaagac gttatcaaag agttcatgcg tttcaaagtt cgtatggaag gttccgttaa
     1081 cggtcacgag ttcgaaatcg aaggtgaagg tgaaggtcgt ccgtacgaag gtacccagac
     1141 cgctaaactg aaagttacca aaggtggtcc gctgccgttc gcttgggaca tcctgtcccc
     1201 gcagttccag tacggttcca aagcttacgt taaacacccg gctgacatcc cggactacct
     1261 gaaactgtcc ttcccggaag gtttcaaatg ggaacgtgtt atgaacttcg aagacggtgg
     1321 tgttgttacc gttacccagg actcctccct gcaagacggt gagttcatct acaaagttaa
     1381 actgcgtggt accaacttcc cgtccgacgg tccggttatg cagaaaaaaa ccatgggttg
     1441 ggaagcttcc accgaacgta tgtacccgga agacggtgct ctgaaaggtg aaatcaaaat
     1501 gcgtctgaaa ctgaaagacg gtggtcacta cgacgctgaa gttaaaacca cctacatggc
     1561 taaaaaaccg gttcagctgc cgggtgctta caaaaccgac atcaaactgg acatcacctc
     1621 ccacaacgaa gactacacca tcgttgaaca gtacgaacgt gctgaaggtc gtcactccac
     1681 cggtgcttaa taacgctgat agtgctagtg tagatcgcta ctagagccag gcatcaaata
     1741 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     1801 gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttataggtct
     1861 catgttgaag agagacgctg cagttgcaag acaatatcga ccgcttgcag cttgataaag
     1921 tgatcaaaga gggacgagca gctacagcac tgctaaggga gatcggagag ccactgccgc
     1981 caccagtgaa agtgaaaaca ggttggttta gtgctgatta ttacggcttt gaggacatgc
     2041 tcacacgttg ggataatgat caatctagcc tttccattta tcacagagag ataaaagagg
     2101 agcaacagga acagcaacga aaacaacagc agatagaaag agaaactaaa caaaaacagg
     2161 aagatgacaa aaagaaacgg caggactatg aaagatattt atcaaaatac ggctatgtca
     2221 aagatgctga gtatgaccat atcagcgacc atgttgcttc agacatttcc atgtttacac
     2281 gtcttcaggc gcaggcatgg gcaagtaacg atatgcaaga gtacataagg tgttcaaagc
     2341 aaaaatacga gcagatcagc gacagaatag cctatgagcg tgacttgaaa aaaatcgggc
     2401 aattgagccg tatcctggat aaagacagac aggcacttgc tgatatttta ccgcagttga
     2461 tgccttacta tgagcagatg caacaggctg ttaataaacg gaaaatttta cttgagaatg
     2521 agcgacagcc atcatttaaa ccacgttatg accaagaaca gcctaagccg aaaaaagaca
     2581 atgacctaac tttttaatgt gttacaatag cattcaaatg taacacattg gagtgaaaaa
     2641 catgacagaa gcgactttta cctttcgggt agaccatgac ctaaagcaag aattttcaag
     2701 ccttgctaag actgttgatc ggtcgggggc gcagctaatc cgtgacttca tgcgtgactt
     2761 tgttaagaag caacaggaag ccgcagacta tgacaaatgg ttcaagcagc aggtacagat
     2821 cggcttaaat gaagccaatg caggcaagtt gattccacat gaggacgtaa aggcagagtt
     2881 tgccgctaga cgtgctgcaa cattggcgaa actggcagcg aagaatgaag attgagtgga
     2941 ctgaaacagc tcgccaagat cgtagaaata tctatgatta tcttgaagaa cgcaacccta
     3001 tagcagctat tgaaattgat gatttaattg aagaaaagac agatttactt gttgataatc
     3061 gactgatggg gcgcacaggc agacagaaag atactaggga gttagtgata catccgcatt
     3121 atgtggttgt atatgacatc actgatataa tacggatact cagagtgcta cacacatcgc
     3181 aggagtggtc atgacttact catgtacttt ggattattta gtgttataaa atcctgattt
     3241 ataaattttt tttgttaaaa aagataaaag ccccttgcaa ttgcttgggg ctttaccgta
     3301 atttatgggg tacagatctt cgatactgac atatcggcaa tcgaaagcat taaggtttga
     3361 cgaccgctaa tgatttcacc acaggggctt aatgtacctg tcttaaattc taaggtttta
     3421 actcgctttg tcaagcatag accccaaaaa tttagccaat gtctgtaact caatctgtcc
     3481 atgtgtgggt gatgaggtac agtgacgcta gcacacatcg gaaaaacgct attactaggg
     3541 gaactgaaca gagtagcgga cgcaatgagt agtcatttaa ttggcggtta tgagcgtgtt
     3601 caggcggtgc tatcaatcgt aatcataaca gtggcagctt gatacagtga tgtcatccct
     3661 gatgcgaaag cgaccgaccg acggtacatc gaatgggaat actttagggt gatttttaag
     3721 aatcgctcta gggtgagtat ttcccattca gctctgctcc ctccctctgg tactttaatc
     3781 aaaagcacta ctaaacatat gtttttaaat aaaaaatatt gatatagaga taatattagt
     3841 aagaataatt aaacaattga atatagataa atcattgtta aataaagatt aattattaaa
     3901 atgaatgtat acttatatat aaatcaatga tttaaaatat ttgataaaga aaacttttca
     3961 aaaaaaatat aattgagatt gtgtcatttc ggtcaattct taatatgttc cacgcaagtt
     4021 ttagctatgg tgctaaacag aaatttgctg aaaaagaact tttcactgaa ctggttaaaa
     4081 tgtaagcagc ctgagagccg ccaaaaattt taaaaacaaa ccgccttaat catcttcaaa
     4141 aaatacctct aaaacctcac catttgcgtt ttaagaccca tatttcatcc tgcccttatg
     4201 ttcccatgct gatagctata aagtgtctgt aatcgcttcc tatgacgttc taggctgttg
     4261 ataacttttg gaacaacgca aaatgttaaa atccgatcat tttttaacct agttattttc
     4321 gttacagggt aactttggta gtattatttc aatatttagt tgttacagga taacaaaata
     4381 tatgttacag ggtaattaat ttagcgttac aggataacta tagttacagg gtaacgttaa
     4441 atagttatcc tgtaatttaa aaatagttgc aggatagcat aaatttttac aaataaattt
     4501 tataaaaagt tatcctgtaa ctaaaaacaa tgttatcctg taactatgat atactatatg
     4561 ggagttaaag acatgaaaga cccgaacgat cagaaaacac aggacatgct aaaagagccg
     4621 ccaaaaacca atgccgaacg acaaaaagca tatcgtgaaa aacgcaaaag ccttgatagt
     4681 caacgtttgg aggtgtttat agataagggt gtgtcggata tgcttgcgga catggtgggc
     4741 gcagcagggg agagccaaaa agccattttg accgcattga ttgagaaaga gtataagcgg
     4801 ctgtatgcag tgaaaaatag caagctaaaa aagtacattg attcaaaaag taatgcagat
     4861 aaaaaagaaa ccctcgattt atgagggttt tttataacta ctaatgccta tgaaaataag
     4921 tgcttatcat tttctctaat gtacccaata aaatcggggt tttgttccac aagtttcttg
     4981 atgtcattat catgtttctt cagtaagtaa gccccgatca ggatcttgcg ttttgcatcg
     5041 tctgaccttg cctgtttttg ctctcgacct ttcaccacag cgtcagctag atcctttctt
     5101 gccttttcct gacgttccaa tttttccttt tcttcatcga gcatttgacc caattccata
     5161 gctgtttttt tgaccatttt taatgcctgt gtctcaacct attttccgaa gtatgaacca
     5221 tcatcaatgt gaaaagtgtt ctacttcaaa aactcgactt acatcaagat ccgagcgcag
     5281 cgagcaaaat caaaaaacaa agtcaaagcc ttattgctct tgcttttgct tttgctttta
     5341 gcctcgcaga gttcccgaag ggcgcactta cgcaaaattt ttgctacgcc aaattttgca
     5401 agtacggtca gggaaacccc gacaccccaa ccgcccaaaa cttggggcgg ctgttaataa
     5461 acaggtgatg aaaatggcaa tttaccattg tgaaatgcag aacatttcga ggtcagatgg
     5521 tcgctcaatc gtggcatgtg cagcataccg agcaggcgaa aaattgtact gtgatacgta
     5581 cggaaaagag caggactaca ccaaaaaaac aggcattgaa tacacccaaa tttttgcccc
     5641 aactggggca agtcctgaca tgttagatcg tcaaacccta tggaatcggg tagagcaatc
     5701 cgaactaaaa aagaacggtg acatcaaaca ggaggcaaga ttagcaaagg aagtagagat
     5761 agcattgccg catgaactgg ataagacaca gcgtcaggca acttgttaac tgcaggcttc
     5821 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5881 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5941 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccacag
     6001 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6061 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6121 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6181 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6241 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6301 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6361 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6421 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6481 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6541 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6601 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6661 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6721 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6781 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6841 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagagccacg
     6901 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6961 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7021 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7081 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7141 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7201 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7261 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7321 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7381 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7441 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7501 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7561 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7621 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7681 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7741 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     7801 gcagaatttc agataaaaaa aatccttagc tttcgctaag gatgatttct ggaattcgcg
     7861 gccgcttcta gagaatagga acttcggaat aggaacttca agatcccctg attccctttg
     7921 tcaacagcaa tggatatcag ccgttgactt agttggtatt tccggaatat catgctgggt
     7981 tccgaataat tttgtttaac tttaagaagg agatatacca tggaagaaca agcgtgaaca
     8041 tggcgcgatt gaagggggag gttttgcccg ccctcttcgc gtcgccggcg acgattggcg
     8101 agtacggggc cggcatcgac ggggcggatt ccctcaacga gctgtcgaat ctgatggagc
     8161 acggcgcagt tgccgcgctg gccgacaaaa tcagccagat cgtggcgaag ctggtcgacg
     8221 cggacccccg caagatcgcg gaaaagccta cctggttcga gaagatgctt ggccgtgagg
     8281 ttgaacgcca ggtgaggtat caggtcgccc gcaagacgct cgaccagttg ctggacgaag
     8341 ccgagggcgt agcgcagcgc gtgcgggaca cgttgcgcgc cttggatgac atgctcaata
     8401 cgcatgaggc cgaggtagac cggctcagag cctacattca agccgggcgc gagttcctgg
     8461 acgagaaccc cgaggccggc gcggccaagg ccggcgtgat cgagttcgac aagccgcgcg
     8521 aacgcttcgc gcgcaagctc gccaacctgg caaccctcat ggcgtcccat gaaatgagcg
     8581 tcactcagat gaagctcacg cgggcgcagg ccgtggacat gctggaccgc ttctctgaaa
     8641 cggcatccgt cctggtgccc gtctggcgtc agcacaccct cgcgctcatc accaccaaga
     8701 acatgaatcc ggcaatggtc gccgaggcgg ccaaagctca ccaggcgctc atgcggagcc
     8761 tttcgcagag cctggaaggc atcaaccaat aacacggcgg gagaacccta tgaacgcact
     8821 gaaaacgacg cacgacgcca aggcccctat cgtcgccttt gacatgaccc cggcaaccct
     8881 gcgcgagctg ggcttgcagg aaagcgacgt gccggaagtc catgcggtcg cgcagcggat
     8941 cgaggtcggc agtccgcaga ccgttgccga gttcggccgc gacgtggccg agcacacgtc
     9001 ccgctacgcc gatagcctgc tggaccaggt gcgcaacagc gacctggacg aagcaggcga
     9061 gaaactgacc caggttgtcg ccaaggcccg ttccctgaac gtcggccctt tgtccgacaa
     9121 ccgttcccgc ctgcccctga ttggcccgct gatcgaccgc tcccgcgtcc gttcgacggg
     9181 cttcatggcg cgcttcgaca cgacccgcga gcagatcgaa cacctggtca gcgaagtgca
     9241 gaccacccag caaggcatcg cgcagcgcaa tgcctcgctc gacgaaatgt tcgcagccgt
     9301 gcgcgaggaa caccgccttc ttggcgtcca catcgcggcc ggcaaggtcc gccttgccga
     9361 gctgcgcgag caggccgagg gtctgcgcgg caatgtcggg aacgaccctg gccgcgtgca
     9421 ggagctggcc gacctcgatg cgatggttgc caacctggac aagcgcatcg gcgacctgat
     9481 cgccttgcaa cattcggcca tgcagagcct gccgaccatc cgcatgatcc aggccaacaa
     9541 ccagatgctg gtcgataaat tccacaccat ccgcgaaatc accgtgccgg cgtggaagcg
     9601 gcaattcatg ctggccttga gcctcaacga gcagaagaac gccgtcgaac tggccacggc
     9661 catcgacgac accaccaacg acctgatgaa gcgcaatgcg gccctgctgc atcgcacgtc
     9721 cgtcgaaacg gcgaaggaga accaacgcct ggtgatcgac gtggacacgc tcaagcaggt
     9781 tcagacgacg ctcatcaaga ccgtcgagga cgttattcgc atccagcagg aaggcgtgca
     9841 gaagcgcaag gatgccgaga agcagatcgc cgcaatgcgt ggcgatcttc aagccaagct
     9901 gacccgccag cccgtgcgcg agctggccca acaggagtcc gtatgaatgc cacaaacacc
     9961 gatgttttcg cccaggtagg cggcctggag gcccgaggcg cgaagatgaa gaagcggggc
    10021 acccgcttcc tcatcgcggc gctggcagtc cttgccattg ccgggatcgg ggcagtaacg
    10081 ggatgggcga tcagcccgag cgcgacgccc ggaagcattg acgtgccgca ggtgctggca
    10141 tcgacattca gcgaccaggt gccgggcagt gagggcggcg gcctgggtgg cggcctgccc
    10201 ttcacttcgg ccgtcggggc attcacggac ttcatggcgg ggccggcaat ttttaccttg
    10261 ggcatt
//
