LOCUS       pJMP12068        6985 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 6985)
  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..6985
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     CDS             complement(264..1124)
                     /codon_start=1
                     /transl_table=11
                     /gene="bla"
                     /product="Bla"
                     /label=bla
                     /db_xref="GI:403391486"
                     /protein_id="AFR43358.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1125..1229)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    1707..6144
                     /label=Tn7 Transposon
     mobile_element  1707..1904
                     /mobile_element_type="transposon:Tn7R"
                     /label=Tn7R
     terminator      1953..2047
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      2150..2236
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    2287..2334
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(2386..3180)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(join(3291..3296,3297..3313,3314..3319))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    3396..3443
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     terminator      complement(3444..3533)
                     /label=spy terminator
     terminator      complement(3622..3672)
                     /label=L3S1P13 terminator
     terminator      complement(3632..3672)
                     /label=L3S1P13 terminator
     terminator      complement(3759..3815)
                     /label=thrLABC terminator
     terminator      complement(3816..3869)
                     /label=garPLRK-rnpB terminator
     CDS             complement(3961..5043)
                     /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"
     promoter        5044..5121
                     /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."
     terminator      5208..5260
                     /label=hisLGDCBHAFI terminator
     terminator      5261..5309
                     /label=pdhR-aceEF-lpd terminator
     misc_feature    join(5386..5392,5393..5408,5409..5414)
                     /label=PabstBR promoter
     misc_feature    5393..5408
                     /label=lac operator
     misc_feature    5416..5445
                     /label=Gibson ends - NcoI/BamHI cloning
     misc_feature    5449..5505
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     misc_feature    5471..5500
                     /label=Gibson ends - NcoI/BamHI cloning
     terminator      5588..5913
                     /label=rrnB terminators
                     /label=rrnB
     terminator      5708..5794
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5886..5913
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     mobile_element  5979..6144
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     misc_feature    6203..6460
                     /label=expanded oriT
     oriT            6216..6324
                     /note="incP origin of transfer"
     rep_origin      6593..6981
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 gcaaaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
       61 acagttacca atgcttaatc agtgaggcac caataactgc ctttgatctt ttctacgggg
      121 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
      181 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
      241 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
      301 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
      361 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgggaccc acgctcaccg
      421 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
      481 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
      541 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
      601 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
      661 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
      721 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
      781 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
      841 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
      901 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
      961 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     1021 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     1081 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     1141 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     1201 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     1261 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     1321 cgtctcgcgc gtttcggtga cgacggtgaa aacctctgac acatgcagct cccggagacg
     1381 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     1441 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     1501 gtgcaccatg tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     1561 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     1621 ctattacgcc agctagagga ccagccgcgt aacctggcaa aatcggttac ggttgagtaa
     1681 taaatggatg ccctgcgtaa gcaggtgtgg gcggacaata aagtcttaaa ctgaacaaaa
     1741 tagatctaaa ctatgacaat aaagtcttaa actagacaga atagttgtaa actgaaatca
     1801 gtccagttat gctgtgaaaa agcatactgg acttttgtta tggctaaagc aaactcttca
     1861 ttttctgaag tgcaaattgc ccgtcgtatt aaagaggggc gtggggttcg aggtcgacgg
     1921 tatcgataag ctagcttaat tagctgagct tggactcctg ttgatagatc cagtaatgac
     1981 ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg
     2041 tgagaatcca agctagactg cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt
     2101 tattggtgcc cttaaacgcc tggggtaatg actctctagc ttgaggcatc aaataaaacg
     2161 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     2221 cctgagtagg acaaatccgc cgctaggagc ttgcggcccg gacgatgagc tcgaattggg
     2281 gatcttgaag tacctattcc gaagttccta ttctctagaa agtataggaa cttcagagcg
     2341 cttttgaagc tgatgctcga gtcatttcga accccagagt cccgctcaga agaactcgtc
     2401 aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag
     2461 gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat
     2521 gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc
     2581 attttccacc atgatattcg gcaagcaggc atcgccgtgg gtcacgacga gatcctcgcc
     2641 gtcgggcatg cgcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc
     2701 ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat
     2761 gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg
     2821 cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc
     2881 ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag
     2941 cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcctg
     3001 cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc
     3061 tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc
     3121 gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat
     3181 gcgaaacgat cctcatcctg tctcttgatc agatcttgat cccctgcgcc atcagatcct
     3241 tggcggcaag aaagccatcc agttttgagc gcatacgcta cttgcattac agtttacgaa
     3301 ccgaacaggc ttatgtcaac tgggttcgtg ccttcatccg tttccacggc tcgagagttc
     3361 aacagacagc cttcgttctt atggccgttc aggaggaagt tcctattccg aagttcctat
     3421 tctctagaaa gtataggaac ttcttgagaa gagaaaagaa aaccgccgat cctgtccacc
     3481 gcattactgc aaggtagtgg acaagaccgg cggtcttaag ttttttggct gaaacgaatg
     3541 acgaaaaggc tgtctgagca aatccaggag gtcgtttaaa ccgtattagc ccggtatttt
     3601 ggaaatagcg gaagcactga cttttgttat caataaaaaa ggccccccgt tagggaggcc
     3661 ttattgttcg tctactcgga agagcgagag acaacagaac ggtcagccac atgaattctt
     3721 tcagctcagt cgataggtag taggcaagag tagtcgcacc tttggtcgaa aaaaaaagcc
     3781 cgcactgtca ggtgcgggct tttttctgtg tttccccaaa agtaaaaacc cgccgaagcg
     3841 ggtttttacg taaaacaggt gaaactgacc agacgagaag gctttggagg tgataatggg
     3901 gctcaaggac ccgggtgggg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     3961 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4021 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     4081 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
     4141 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     4201 acatgagcta tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag
     4261 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     4321 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     4381 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
     4441 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
     4501 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccat cttcatggga
     4561 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
     4621 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
     4681 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
     4741 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
     4801 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
     4861 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     4921 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     4981 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     5041 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     5101 tttgcaccat tcgatggtgt caacgtaaat gcatgccgct tccccgactc ggtgccactt
     5161 tttcgggagg cagataggta ggcatggccc ccattttcaa tacaagcaac gcatgagaaa
     5221 gcccccggaa gatcaccttc cgggggcttt tttattgcgc tccggcaatt aaaaaagcgg
     5281 ctaaccacgc cgcttttttt acgtctgcag actagtcagg cagccatcgg aagctgtggt
     5341 atggctgtgc aggtcgtaaa gacgtctcat taatcagata aaatattgac aatgtgagcg
     5401 gataacaaga tactgagcac atcagcagca cacaggaaac agaccatgga attcgagctc
     5461 ggtacccggg gatcctctag agtcgacctg caggcatgca agcttggctg ttttggcgga
     5521 tgagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa
     5581 cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa
     5641 gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc
     5701 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     5761 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc
     5821 gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat
     5881 taagcagaag gccatcctga cggatggcct tttcacagaa tgagcatcat ggcctagtcg
     5941 acatcggaga agagtacggc tcttttaacc gcctcaagaa ccagataagt gaaatctagt
     6001 tccaaactat tttgtcattt ttaattttcg tattagctta cgacgctaca cccagttccc
     6061 atctattttg tcactcttcc ctaaataatc cttaaaaact ccatttccac ccctcccagt
     6121 tcccaactat tttgtccgcc cacagcgggg catttttctt cctgttatgt ttgggctctt
     6181 ccgcttcctc gctcactgac cctatcagag cttatcggcc agcctcgcag agcaggattc
     6241 ccgttgagca ccgccaggtg cgaataaggg acagtgaaga aggaacaccc gctcgcgggt
     6301 gggcctactt cacctatcct gcccggctga cgccgttgga tacaccaagg aaagtctaca
     6361 cgaacccttt ggcaaaatcc tgtatatcgt gcgaaaaagg atggatatac cgaaaaaatc
     6421 gctataatga ccccgaagca gggttatgca gcggaaagta taccttaagg aatcccctga
     6481 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6541 cgcactccct gcctctgtca tcacgatact gtgatgccgt ggctaattcc catgtcagcc
     6601 gttaagtgtt cctgtgtcac tcaaaattgc tttgagaggc tctaagggct tctcagtgcg
     6661 ttacatccct ggcttgttgt ccacaaccgt taaaccttaa aagctttaaa agccttatat
     6721 attctttttt ttcttataaa acttaaaacc ttagaggcta tttaagttgc tgatttatat
     6781 taattttatt gttcaaacat gagagcttag tacgtgaaac atgagagctt agtacgttag
     6841 ccatgagagc ttagtacgtt agccatgagg gtttagttcg ttaaacatga gagcttagta
     6901 cgttaaacat gagagcttag tacgtgaaac atgagagctt agtacgtact atcaacaggt
     6961 tgaactgctg atcttcagat cggcc
//
