LOCUS       pJMP3666        6411 bp DNA     circular SYN 30-MAY-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6411)
  AUTHORS   Jason Peters Lab
  TITLE     Direct Submission
  JOURNAL   Exported May 30, 2024 from SnapGene 7.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6411
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     promoter        27..104
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             105..1187
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     misc_feature    join(1344..1350,1351..1366,1367..1372)
                     /label=PabstBR promoter
     misc_feature    1351..1366
                     /label=lac operator
     misc_feature    1392..1402
                     /label=strong RBS
     CDS             1404..2117
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     misc_feature    2122..2152
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      2235..2560
                     /label=rrnB terminators
                     /label=rrnB
     terminator      2355..2441
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2533..2560
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        join(2676..2681,2682..2698,2699..2704)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2893..3693
                     /codon_start=1
                     /gene="aac(3)IV"
                     /product="aminoglycoside 3-N-acetyltransferase type IV     
                        (apramycin resistance)       
                     
                     is from Addgene121998 (pJMP736) but with restriction enzyme
                     sites removed"
                     /label=AprR
                     /note="confers resistance to apramycin (Paget and Davies, 
                     1996)"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKENSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     misc_feature    3789..5130
                     /label=A. baumannii pWH1266 origin
                     /note="from pSGA-km"
     misc_feature    5189..5424
                     /label=expanded oriT
     oriT            5315..5424
                     /note="incP origin of transfer"
     rep_origin      5807..6352
                     /direction=RIGHT
                     /label=E. coli p15A origin
                     /note="medium-copy-number p15A origin of replication can be
                     propagated in E. coli"
ORIGIN
        1 aaggcgaagc ggcatgcatt tacgttgaca ccatcgaatg gtgcaaaacc tttcgcggta
       61 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt
      121 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
      181 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
      241 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
      301 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
      361 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
      421 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
      481 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
      541 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg
      601 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
      661 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
      721 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
      781 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
      841 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
      901 cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt
      961 taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
     1021 aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa
     1081 gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     1141 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     1201 aatgtgagtt agcgcgaatt gatctggttt gacagcttat catcgactgc acggtgcacc
     1261 aatgcttctg gcgtcaggca gccatcggaa gctgtggtat ggctgtgcag gtcgtaaaga
     1321 cgtctcatta atcagataaa atattgacaa tgtgagcgga taacaagata ctgagcacat
     1381 cagcagcaca caggaaacag accatgagca aaggagaaga acttttcact ggagttgtcc
     1441 caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc cgtggagagg
     1501 gtgaaggtga tgctacaaac ggaaaactca cccttaaatt tatttgcact actggaaaac
     1561 tacctgttcc gtggccaaca cttgtcacta ctctgaccta tggtgttcaa tgcttttccc
     1621 gttatccgga tcacatgaaa cggcatgact ttttcaagag tgccatgccc gaaggttatg
     1681 tacaggaacg cactatatct ttcaaagatg acgggaccta caagacgcgt gctgaagtca
     1741 agtttgaagg tgataccctt gttaatcgta tcgagttaaa gggtattgat tttaaagaag
     1801 atggaaacat tcttggacac aaactcgagt acaactttaa ctcacacaat gtatacatca
     1861 cggcagacaa acaaaagaat ggaatcaaag ctaacttcaa aattcgccac aacgttgaag
     1921 atggttccgt tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg
     1981 tccttttacc agacaaccat tacctgtcga cacaatctgt cctttcgaaa gatcccaacg
     2041 aaaagcgtga ccacatggtc cttcttgagt ttgtaactgc tgctgggatt acacatggca
     2101 tggatgagct ctacaaagat cctctagagt cgacctgcag gcatgcaagc ttggctgttt
     2161 tggcggatga gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct
     2221 gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa
     2281 ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg
     2341 gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     2401 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
     2461 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc
     2521 atcaaattaa gcagaaggcc atcctgacgg atggcctttt tcctgaacgg ccataagaac
     2581 gaaggctgtc tgttgaactc tcgagccttg tcgccttgcg tataatattt gctcatggac
     2641 gcacaccgtg gaaacggatg aaggcacgaa cccagttgac ataagcctgt tcggttcgta
     2701 aactgtaatg caagtagcgt atgcgctcac gcaactggtc cagaaccttg accgaacgca
     2761 gcggtggtaa cggcgcagtg gcggttttca tggcttgtta tgactgtttt ttcgtacagt
     2821 ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat
     2881 ggagcagcaa cgatgtcatc agcggtggag tgcaatgtcg tgcaatacga atggcgaaaa
     2941 gccgagctga tcggtcagct tctcaacctt ggggttaccc ccggcggtgt gctgctggtc
     3001 cacagctcct tccgtagcgt ccggcccctc gaagatgggc cacttggact gatcgaggcc
     3061 ctgcgtgctg cactgggtcc gggagggacg ctcgtcatgc catcgtggtc aggtctggac
     3121 gacgagccgt tcgatcctgc cacgtcgccc gttacaccgg accttggagt tgtctctgac
     3181 acattctggc gtctgccaaa tgtaaagcgc agcgcccatc catttgcctt tgcggcagcg
     3241 gggccacagg cagagcagat catctctgat ccattgcccc tgccacctca ctcgcctgca
     3301 agcccggtcg cccgtgtcca tgaactcgat gggcaggtac ttctcctcgg cgtgggacac
     3361 gatgccaaca cgacgctgca tcttgccgag ttgatggcaa aggttcccta tggggtgccg
     3421 agacactgca ccattcttca ggatggcaag ttggtacgtg tcgattatct ggagaatgac
     3481 cactgctgtg agcgctttgc cttggcggac aggtggctca aggagaacag ccttcagaag
     3541 gaaggtccag tcggtcatgc ctttgctcgg ttgatccgct cccgcgacat tgtggcgaca
     3601 gccctgggtc aactgggccg agatccgttg atcttcctgc atccgccaga ggcgggatgc
     3661 gaagaatgcg atgccgctcg ccagtcgatt ggctaacaat tcgttcaagc cgagatctcg
     3721 agcatcagct tcaaaagcgc tccgatgtac tgcggtttct ggactagtcc gttgacatta
     3781 tcgcagcagg attttaacat tttgcgttgt tccaaaagtt atcaacagcc tagaacgtca
     3841 taggaagcga ttacagacac tttatagcta tcagcatggg aacataaggg caggatgaaa
     3901 tatgggtctt aaaacgcaaa tggtgaggtt ttagaggtat tttttgaaga tgattaaggc
     3961 ggtttgtttt taaaattttt ggcggctctc aggctgctta cattttaacc agttcagtga
     4021 aaagttcttt ttcagcaaat ttctgtttag caccatagct aaaacttgcg tggaacatat
     4081 taagaattga ccgaaatgac acaatctcaa ttatattttt tttgaaaagt tttctttatc
     4141 aaatatttta aatcattgat ttatatataa gtatgcattc attttaataa ttaatcttta
     4201 tttaacaatg atttatctat attcaattgt ttaattattc ttactaatat tatctctata
     4261 tcaatatttt ttatttaaaa acatatgttt agtagtgctt ttgattaaag taccagaggg
     4321 agggagcaga gctgaatggg aaatactcac cctagagcga ttcttaaaaa tcaccctaaa
     4381 gtattcccat tcgatgtacc gtcggtcggt cgctttcgca tcagggatga catcactgta
     4441 tcaagctgcc actgttatga ttacgattga tagcaccgcc tgaacacgct cataaccgcc
     4501 aattaaatga ctactcattg cgtccgctac tctgttcagt tcccctagta atagcgtttt
     4561 tccgatgtgt gctagcgtca ctgtacctca tcacccacac atggacagat tgagttacag
     4621 acattggcta aatttttggg gtctatgctt gacaaagcga gttaaaacct tagaatttaa
     4681 gacaggtaca ttaagcccct gtggtgaaat cattagcggt cgtcaaacct taatgctttc
     4741 gattgccgat atgtcagtat cgaagatctg taccccataa attacggtaa agccccaagc
     4801 aattgcaagg ggcttttatc ttttttaaca aaaaaaattt ataaatcagg attttataac
     4861 actaaataat ccaaagtaca tgagtaagtc atgaccactc ctgcgatgtg tgtagcactc
     4921 tgagtatccg tattatatca gtgatgtcat atacaaccac ataatgcgga tgtatcacta
     4981 actccctagt atctttctgt ctgcctgtgc gccccatcag tcgattatca acaagtaaat
     5041 ctgtcttttc ttcaattaaa tcatcaattt caatagctgc tatagggttg cgttcttcaa
     5101 gataatcata gatatttcta cgatcttggc ttgcagccct tatgtcctca ggcgcgcaca
     5161 aaacagcagg gaagcagcgc ttttccgctg cataaccctg cttcggggtc attatagcga
     5221 ttttttcggt atatccatcc tttttcgcac gatatacagg attttgccaa agggttcgtg
     5281 tagactttcc ttggtgtatc caacggcgtc agccgggcag gataggtgaa gtaggcccac
     5341 ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca cctggcggtg ctcaacggga
     5401 atcctgctct gcgaggctgg ccggctaccg ccggcgtaac agatgagggc aagcggatgg
     5461 ctgatgaaac caagccaacc aggaagggca gcccacctat caaggtgtac tgccttccag
     5521 acgaacgaag agcgattgag gaaaaggcgg cggcggccgg catgagcctg tcggcctacc
     5581 tgctggccgt cggccagggc tacaaaatca cgggcgtcgt ggactatgag cacgtccgcg
     5641 agctggcccg catcaatggc gacctgggcc gcctgggcgg cctgctgaaa ctctggctca
     5701 ccgacgaccc gcgcacggcg cggttcggtg atgccacgat cctcgccctg ctggcgaaga
     5761 tcgactctag ctagaaatat tttatctgat taataagatg atcttcttga gatcgttttg
     5821 gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc ggtttttcga
     5881 aggttctctg agctaccaac tctttgaacc gaggtaactg gcttggagga gcgcagtcac
     5941 caaaacttgt cctttcagtt tagccttaac cggcgcatga cttcaagact aactcctcta
     6001 aatcaattac cagtggctgc tgccagtggt gcttttgcat gtctttccgg gttggactca
     6061 agacgatagt taccggataa ggcgcagcgg tcggactgaa cggggggttc gtgcatacag
     6121 tccagcttgg agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg
     6181 cggccataac agcggaatga caccggtaaa ccgaaaggca ggaacaggag agcgcacgag
     6241 ggagccgcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccaccactg
     6301 atttgagcgt cagatttcgt gatgcttgtc aggggggcgg agcctatgga aaaacggctt
     6361 tgccgcggcc ctctcacttc cctgttaaac cggaattcta ggcccaaacg g
//
