LOCUS       sequence-404864-       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-E-TelR
FEATURES             Location/Qualifiers
     source          1..10260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    100..121
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(136..141,142..159,160..166)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    174..190
                     /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             208..219
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             226..903
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      948..1019
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1035..1062
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(5028..5045)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5199..5787)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5279..5298)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5958..6749,6750..6818))
                     /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     6581..6600
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6819..6923)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6991..7009
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(7013..7056)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    7059..7078
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     terminator      7158..7189
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     RBS             10090..10112
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
       61 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
      121 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
      181 cggataacaa tttcacacat actagagaaa gaggagaaat actagatggc ttcctccgaa
      241 gacgttatca aagagttcat gcgtttcaaa gttcgtatgg aaggttccgt taacggtcac
      301 gagttcgaaa tcgaaggtga aggtgaaggt cgtccgtacg aaggtaccca gaccgctaaa
      361 ctgaaagtta ccaaaggtgg tccgctgccg ttcgcttggg acatcctgtc cccgcagttc
      421 cagtacggtt ccaaagctta cgttaaacac ccggctgaca tcccggacta cctgaaactg
      481 tccttcccgg aaggtttcaa atgggaacgt gttatgaact tcgaagacgg tggtgttgtt
      541 accgttaccc aggactcctc cctgcaagac ggtgagttca tctacaaagt taaactgcgt
      601 ggtaccaact tcccgtccga cggtccggtt atgcagaaaa aaaccatggg ttgggaagct
      661 tccaccgaac gtatgtaccc ggaagacggt gctctgaaag gtgaaatcaa aatgcgtctg
      721 aaactgaaag acggtggtca ctacgacgct gaagttaaaa ccacctacat ggctaaaaaa
      781 ccggttcagc tgccgggtgc ttacaaaacc gacatcaaac tggacatcac ctcccacaac
      841 gaagactaca ccatcgttga acagtacgaa cgtgctgaag gtcgtcactc caccggtgct
      901 taataacgct gatagtgcta gtgtagatcg ctactagagc caggcatcaa ataaaacgaa
      961 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct
     1021 actagagtca cactggctca ccttcgggtg ggcctttctg cgtttatagg tctcatgttt
     1081 tagagagacg ctgcagttaa caagttgcct gacgctgtgt cttatccagt tcatgcggca
     1141 atgctatctc tacttccttt gctaatcttg cctcctgttt gatgtcaccg ttctttttta
     1201 gttcggattg ctctacccga ttccataggg tttgacgatc taacatgtca ggacttgccc
     1261 cagttggggc aaaaatttgg gtgtattcaa tgcctgtttt tttggtgtag tcctgctctt
     1321 ttccgtacgt atcacagtac aatttttcgc ctgctcggta tgctgcacat gccacgattg
     1381 agcgaccatc tgacctcgaa atgttctgca tttcacaatg gtaaattgcc attttcatca
     1441 cctgtttatt aacagccgcc ccaagttttg ggcggttggg gtgtcggggt ttccctgacc
     1501 gtacttgcaa aatttggcgt agcaaaaatt ttgcgtaagt gcgcccttcg ggaactctgc
     1561 gaggctaaaa gcaaaagcaa gagcaataag gctttgactt tgttttttga ttttgctcgc
     1621 tgcgctcgga tcttgatgta agtcgagttt ttgaagtaga acacttttca cattgatgat
     1681 ggttcatact tcggaaaata ggttgagaca caggcattaa aaatggtcaa aaaaacagct
     1741 atggaattgg gtcaaatgct cgatgaagaa aaggaaaaat tggaacgtca ggaaaaggca
     1801 agaaaggatc tagctgacgc tgtggtgaaa ggtcgagagc aaaaacaggc aaggtcagac
     1861 gatgcaaaac gcaagatcct gatcggggct tacttactga agaaacatga taatgacatc
     1921 aagaaacttg tggaacaaaa ccccgatttt attgggtaca ttagagaaaa tgataagcac
     1981 ttattttcat aggcattagt agttataaaa aaccctcata aatcgagggt ttctttttta
     2041 tctgcattac tttttgaatc aatgtacttt tttagcttgc tatttttcac tgcatacagc
     2101 cgcttatact ctttctcaat caatgcggtc aaaatggctt tttggctctc ccctgctgcg
     2161 cccaccatgt ccgcaagcat atccgacaca cccttatcta taaacacctc caaacgttga
     2221 ctatcaaggc ttttgcgttt ttcacgatat gctttttgtc gttcggcatt ggtttttggc
     2281 ggctctttta gcatgtcctg tgttttctga tcgttcgggt ctttcatgtc tttaactccc
     2341 atatagtata tcatagttac aggataacat tgtttttagt tacaggataa ctttttataa
     2401 aatttatttg taaaaattta tgctatcctg caactatttt taaattacag gataactatt
     2461 taacgttacc ctgtaactat agttatcctg taacgctaaa ttaattaccc tgtaacatat
     2521 attttgttat cctgtaacaa ctaaatattg aaataatact accaaagtta ccctgtaacg
     2581 aaaataacta ggttaaaaaa tgatcggatt ttaacatttt gcgttgttcc aaaagttatc
     2641 aacagcctag aacgtcatag gaagcgatta cagacacttt atagctatca gcatgggaac
     2701 ataagggcag gatgaaatat gggtcttaaa acgcaaatgg tgaggtttta gaggtatttt
     2761 ttgaagatga ttaaggcggt ttgtttttaa aatttttggc ggctctcagg ctgcttacat
     2821 tttaaccagt tcagtgaaaa gttctttttc agcaaatttc tgtttagcac catagctaaa
     2881 acttgcgtgg aacatattaa gaattgaccg aaatgacaca atctcaatta tatttttttt
     2941 gaaaagtttt ctttatcaaa tattttaaat cattgattta tatataagta tacattcatt
     3001 ttaataatta atctttattt aacaatgatt tatctatatt caattgttta attattctta
     3061 ctaatattat ctctatatca atatttttta tttaaaaaca tatgtttagt agtgcttttg
     3121 attaaagtac cagagggagg gagcagagct gaatgggaaa tactcaccct agagcgattc
     3181 ttaaaaatca ccctaaagta ttcccattcg atgtaccgtc ggtcggtcgc tttcgcatca
     3241 gggatgacat cactgtatca agctgccact gttatgatta cgattgatag caccgcctga
     3301 acacgctcat aaccgccaat taaatgacta ctcattgcgt ccgctactct gttcagttcc
     3361 cctagtaata gcgtttttcc gatgtgtgct agcgtcactg tacctcatca cccacacatg
     3421 gacagattga gttacagaca ttggctaaat ttttggggtc tatgcttgac aaagcgagtt
     3481 aaaaccttag aatttaagac aggtacatta agcccctgtg gtgaaatcat tagcggtcgt
     3541 caaaccttaa tgctttcgat tgccgatatg tcagtatcga agatctgtac cccataaatt
     3601 acggtaaagc cccaagcaat tgcaaggggc ttttatcttt tttaacaaaa aaaatttata
     3661 aatcaggatt ttataacact aaataatcca aagtacatga gtaagtcatg accactcctg
     3721 cgatgtgtgt agcactctga gtatccgtat tatatcagtg atgtcatata caaccacata
     3781 atgcggatgt atcactaact ccctagtatc tttctgtctg cctgtgcgcc ccatcagtcg
     3841 attatcaaca agtaaatctg tcttttcttc aattaaatca tcaatttcaa tagctgctat
     3901 agggttgcgt tcttcaagat aatcatagat atttctacga tcttggcgag ctgtttcagt
     3961 ccactcaatc ttcattcttc gctgccagtt tcgccaatgt tgcagcacgt ctagcggcaa
     4021 actctgcctt tacgtcctca tgtggaatca acttgcctgc attggcttca tttaagccga
     4081 tctgtacctg ctgcttgaac catttgtcat agtctgcggc ttcctgttgc ttcttaacaa
     4141 agtcacgcat gaagtcacgg attagctgcg cccccgaccg atcaacagtc ttagcaaggc
     4201 ttgaaaattc ttgctttagg tcatggtcta cccgaaaggt aaaagtcgct tctgtcatgt
     4261 ttttcactcc aatgtgttac atttgaatgc tattgtaaca cattaaaaag ttaggtcatt
     4321 gtcttttttc ggcttaggct gttcttggtc ataacgtggt ttaaatgatg gctgtcgctc
     4381 attctcaagt aaaattttcc gtttattaac agcctgttgc atctgctcat agtaaggcat
     4441 caactgcggt aaaatatcag caagtgcctg tctgtcttta tccaggatac ggctcaattg
     4501 cccgattttt ttcaagtcac gctcataggc tattctgtcg ctgatctgct cgtatttttg
     4561 ctttgaacac cttatgtact cttgcatatc gttacttgcc catgcctgcg cctgaagacg
     4621 tgtaaacatg gaaatgtctg aagcaacatg gtcgctgata tggtcatact cagcatcttt
     4681 gacatagccg tattttgata aatatctttc atagtcctgc cgtttctttt tgtcatcttc
     4741 ctgtttttgt ttagtttctc tttctatctg ctgttgtttt cgttgctgtt cctgttgctc
     4801 ctcttttatc tctctgtgat aaatggaaag gctagattga tcattatccc aacgtgtgag
     4861 catgtcctca aagccgtaat aatcagcact aaaccaacct gttttcactt tcactggtgg
     4921 cggcagtggc tctccgatct cccttagcag tgctgtagct gctcgtccct ctttgatcac
     4981 tttatcaagc tgcaagcggt cgatattgtc ttgcaactgc aggcttcctc gctcactgac
     5041 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5101 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5161 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccacaggct ccgcccccct
     5221 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5281 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5341 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5401 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5461 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5521 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5581 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5641 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5701 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5761 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5821 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5881 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5941 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6001 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6061 ggcttaccat ctggccccag tgctgcaatg ataccgcgag agccacgctc accggctcca
     6121 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6181 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6241 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6301 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6361 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6421 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6481 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6541 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6601 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6661 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6721 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6781 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6841 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6901 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     6961 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggca gaatttcaga
     7021 taaaaaaaat ccttagcttt cgctaaggat gatttctgga attcgcggcc gcttctagaa
     7081 agtataggaa cttcagagcg cttttgaagc tgatgtgctt aaaaacttac tcaatggaat
     7141 tagctcctcg agcagaaagt caaaagcctc cgaccggagg cttttgactt gagggggatc
     7201 gatcccttat ggctctgcac ccggctccat caccaacagg tcgcgcacgc gcttcactcg
     7261 gttgcggatc gacactgcca gcccaacaaa gccggttgcc gccgccgcca ggatcgcgcc
     7321 gatgatgccg gccacaccgg ccatcgccca ccaggtcgcc gccttccggt tccattcctg
     7381 ctggtactgc ttcgcaatgc tggacctcgg ctcaccatag gctgaccgct cgatggcgta
     7441 tgccgcttct ccccttggcg taaaacccag cgccgcaggc ggcattgcca tgctgcccgc
     7501 cgctttcccg accacgacgc gcgcaccagg cttgcggtcc agaccttcgg ccacggcgag
     7561 ctgcgcaagg acataatcag ccgccgactt ggctccacgc gcctcgatca gctcttgcac
     7621 tcgcgcgaaa tccttggcct ccacggccgc catgaatcgc gcacgcggcg aaggctccgc
     7681 agggccggcg tcgtgatcgc cgccgagaat gcccttcacc aagttcgacg acacgaaaat
     7741 catgctgacg gctatcacca tcatgcagac ggatcgcacg aacccgcaga actcaccccc
     7801 gaacacgagc acggcacccg cgaccactat gccaagaatg cccaaggtaa aaattgccgg
     7861 ccccgccatg aagtccgtga atgccccgac ggccgaagtg aagggcaggc cgccacccag
     7921 gccgccgccc tcactgcccg gcacctggtc gctgaatgtc gatgccagca cctgcggcac
     7981 gtcaatgctt ccgggcgtcg cgctcgggct gatcgcccat cccgttactg ccccgatccc
     8041 ggcaatggca aggactgcca gcgccgcgat gaggaagcgg gtgccccgct tcttcatctt
     8101 cgcgcctcgg gcctccaggc cgcctacctg ggcgaaaaca tcggtgtttg tggcattcat
     8161 acggactcct gttgggccag ctcgcgcacg ggctggcggg tcagcttggc ttgaagatcg
     8221 ccacgcattg cggcgatctg cttctcggca tccttgcgct tctgcacgcc ttcctgctgg
     8281 atgcgaataa cgtcctcgac ggtcttgatg agcgtcgtct gaacctgctt gagcgtgtcc
     8341 acgtcgatca ccaggcgttg gttctccttc gccgtttcga cggacgtgcg atgcagcagg
     8401 gccgcattgc gcttcatcag gtcgttggtg gtgtcgtcga tggccgtggc cagttcgacg
     8461 gcgttcttct gctcgttgag gctcaaggcc agcatgaatt gccgcttcca cgccggcacg
     8521 gtgatttcgc ggatggtgtg gaatttatcg accagcatct ggttgttggc ctggatcatg
     8581 cggatggtcg gcaggctctg catggccgaa tgttgcaagg cgatcaggtc gccgatgcgc
     8641 ttgtccaggt tggcaaccat cgcatcgagg tcggccagct cctgcacgcg gccagggtcg
     8701 ttcccgacat tgccgcgcag accctcggcc tgctcgcgca gctcggcaag gcggaccttg
     8761 ccggccgcga tgtggacgcc aagaaggcgg tgttcctcgc gcacggctgc gaacatttcg
     8821 tcgagcgagg cattgcgctg cgcgatgcct tgctgggtgg tctgcacttc gctgaccagg
     8881 tgttcgatct gctcgcgggt cgtgtcgaag cgcgccatga agcccgtcga acggacgcgg
     8941 gagcggtcga tcagcgggcc aatcaggggc aggcgggaac ggttgtcgga caaagggccg
     9001 acgttcaggg aacgggcctt ggcgacaacc tgggtcagtt tctcgcctgc ttcgtccagg
     9061 tcgctgttgc gcacctggtc cagcaggcta tcggcgtagc gggacgtgtg ctcggccacg
     9121 tcgcggccga actcggcaac ggtctgcgga ctgccgacct cgatccgctg cgcgaccgca
     9181 tggacttccg gcacgtcgct ttcctgcaag cccagctcgc gcagggttgc cggggtcatg
     9241 tcaaaggcga cgataggggc cttggcgtcg tgcgtcgttt tcagtgcgtt catagggttc
     9301 tcccgccgtg ttattggttg atgccttcca ggctctgcga aaggctccgc atgagcgcct
     9361 ggtgagcttt ggccgcctcg gcgaccattg ccggattcat gttcttggtg gtgatgagcg
     9421 cgagggtgtg ctgacgccag acgggcacca ggacggatgc cgtttcagag aagcggtcca
     9481 gcatgtccac ggcctgcgcc cgcgtgagct tcatctgagt gacgctcatt tcatgggacg
     9541 ccatgagggt tgccaggttg gcgagcttgc gcgcgaagcg ttcgcgcggc ttgtcgaact
     9601 cgatcacgcc ggccttggcc gcgccggcct cggggttctc gtccaggaac tcgcgcccgg
     9661 cttgaatgta ggctctgagc cggtctacct cggcctcatg cgtattgagc atgtcatcca
     9721 aggcgcgcaa cgtgtcccgc acgcgctgcg ctacgccctc ggcttcgtcc agcaactggt
     9781 cgagcgtctt gcgggcgacc tgatacctca cctggcgttc aacctcacgg ccaagcatct
     9841 tctcgaacca ggtaggcttt tccgcgatct tgcgggggtc cgcgtcgacc agcttcgcca
     9901 cgatctggct gattttgtcg gccagcgcgg caactgcgcc gtgctccatc agattcgaca
     9961 gctcgttgag ggaatccgcc ccgtcgatgc cggccccgta ctcgccaatc gtcgccggcg
    10021 acgcgaagag ggcgggcaaa acctccccct tcaatcgcgc catgttcacg cttgttcttc
    10081 catggtatat ctccttctta aagttaaaca aaattattcg gaacccagca tgatattccg
    10141 gaaataccaa ctaagtcaac ggctgatatc cattgctgtt gacaaaggga atcaggggat
    10201 cttgaagttc ctattccgaa gttcctattc tctagaacgt ctcacttcct atagagaccc
//
