LOCUS       pJMP12089        7306 bp DNA     circular SYN 30-MAY-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7306)
  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..7306
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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..6465
                     /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(2385..3383)
                     /codon_start=1
                     /product="hygromycin resistance gene from Streptomyces 
                     hygroscopicus optimized for Ecoli and Mobile CRIPSRi 
                     restriction sites removed
                     "
                     /label=hygR
                     /translation="MTQESLLLLDRIDSDDSYASLRNDQEFWEPLARRALEELGLPVPP
                     VLRVPGESTNPVLVGEPDPVIKLFGEHWCGPESLASESEAYAVLADAPVPVPRLLGRGE
                     LRPGTGAWPWPYLVMSRMTGTTWRSAMDGTTDRNALLALARELGRVLGRLHRVPLTGNT
                     VLTPHSEVFPELLRERRAATVEDHRGWGYLSPRLLDRLEDWLPDVDTLLAGREPRFVHG
                     DLHGTNIFVDLAATEVTGIVDFTDVYAGDSRYSLVQLHLNAFRGDREILAALLDGAQWK
                     RTEDFARELLAFTFLHDFEVFEETPLDLSGFTDPEELAQFLWGPPDTAPGA"
     promoter        complement(join(3572..3577,3578..3594,3595..3600))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    3717..3764
                     /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(3765..3854)
                     /label=spy terminator
     terminator      complement(3943..3993)
                     /label=L3S1P13 terminator
     terminator      complement(3953..3993)
                     /label=L3S1P13 terminator
     terminator      complement(4080..4136)
                     /label=thrLABC terminator
     terminator      complement(4137..4190)
                     /label=garPLRK-rnpB terminator
     CDS             complement(4282..5364)
                     /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        5365..5442
                     /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      5529..5581
                     /label=hisLGDCBHAFI terminator
     terminator      5582..5630
                     /label=pdhR-aceEF-lpd terminator
     misc_feature    join(5707..5713,5714..5729,5730..5735)
                     /label=PabstBR promoter
     misc_feature    5714..5729
                     /label=lac operator
     misc_feature    5737..5766
                     /label=Gibson ends - NcoI/BamHI cloning
     misc_feature    5770..5826
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     misc_feature    5792..5821
                     /label=Gibson ends - NcoI/BamHI cloning
     terminator      5909..6234
                     /label=rrnB terminators
                     /label=rrnB
     terminator      6029..6115
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6207..6234
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     mobile_element  6300..6465
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     misc_feature    6524..6781
                     /label=expanded oriT
     oriT            6537..6645
                     /note="incP origin of transfer"
     rep_origin      6914..7302
                     /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 gatctcggct tgaacgaatt gttattatgc acccggtgcg
     2401 gtatcaggcg gaccccacag aaactgtgct aattcttccg gatcggtaaa accgctcaga
     2461 tccagcggtg tttcttcaaa cacttcaaaa tcatgcagaa aggtaaaggc cagcagttca
     2521 cgggcaaaat cttcggtacg tttccactgt gcaccatcca gcagggctgc cagaatttca
     2581 cgatcaccac gaaatgcatt cagatgcagc tgaaccaggc tataacggct atcaccggca
     2641 taaacatcgg tgaaatcaac aataccggta acttcggttg ctgccagatc aacaaaaatg
     2701 ttggtgccat gcagatcacc atgaacaaaa cgcggttcgc gacctgccag cagggtatca
     2761 acatccggca gccaatcttc cagacggtcc agcagacgcg gactcagata accccaacca
     2821 cgatgatctt caacggttgc ggcacgacgt tcacgcagca gttccggaaa aacttcgcta
     2881 tgcggtgtca gaacggtatt accggtcagc ggaacacgat gcagacgacc cagaacacgt
     2941 cccagttcac gtgccagtgc cagcagtgca ttacgatcgg tggtgccatc cattgcgcta
     3001 cgccaggtgg tgccggtcat acggctcata accagatacg gccacggcca tgcacctgta
     3061 ccaggacgca gttcaccacg acctaacaga cgaggaaccg gaaccggtgc atctgccaga
     3121 actgcatagg cttcgctttc gcttgccagg ctttccggac cacaccaatg ttcaccaaac
     3181 agtttaataa caggatcagg ttcaccaacc agaaccggat tggtgctttc acccggaaca
     3241 cgcagaacag gcggaaccgg cagacccagt tcttccagtg cacgacgtgc cagcggttcc
     3301 caaaattctt gatcattgcg caggcttgca tagctatcat cactatcaat acgatccagc
     3361 agcagcaggc tttcttgggt catcgttgct gctccataac atcaaacatc gacccacggc
     3421 gtaacgcgct tgctgcttgg atgcccgagg catagactgt acgaaaaaac agtcataaca
     3481 agccatgaaa accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca
     3541 gttgcgtgag cgcatacgct acttgcatta cagtttacga accgaacagg cttatgtcaa
     3601 ctgggttcgt gccttcatcc gtttccacgg tgtgcgtcca tgagcaaata ttatacgcaa
     3661 ggcgacaagg ctcgagagtt caacagacag ccttcgttct tatggccgtt caggaggaag
     3721 ttcctattcc gaagttccta ttctctagaa agtataggaa cttcttgaga agagaaaaga
     3781 aaaccgccga tcctgtccac cgcattactg caaggtagtg gacaagaccg gcggtcttaa
     3841 gttttttggc tgaaacgaat gacgaaaagg ctgtctgagc aaatccagga ggtcgtttaa
     3901 accgtattag cccggtattt tggaaatagc ggaagcactg acttttgtta tcaataaaaa
     3961 aggccccccg ttagggaggc cttattgttc gtctactcgg aagagcgaga gacaacagaa
     4021 cggtcagcca catgaattct ttcagctcag tcgataggta gtaggcaaga gtagtcgcac
     4081 ctttggtcga aaaaaaaagc ccgcactgtc aggtgcgggc ttttttctgt gtttccccaa
     4141 aagtaaaaac ccgccgaagc gggtttttac gtaaaacagg tgaaactgac cagacgagaa
     4201 ggctttggag gtgataatgg ggctcaagga cccgggtggg gtgcctaatg agtgagctaa
     4261 cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4321 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt
     4381 ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg
     4441 agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat
     4501 ggtggttaac ggcgggatat aacatgagct atcttcggta tcgtcgtatc ccactaccga
     4561 gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat
     4621 ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt
     4681 ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg
     4741 attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa
     4801 tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag
     4861 tcgcgtacca tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc
     4921 aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc
     4981 cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc
     5041 tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg
     5101 atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga
     5161 ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg
     5221 aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg
     5281 gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac
     5341 atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta
     5401 tcatgccata ccgcgaaagg ttttgcacca ttcgatggtg tcaacgtaaa tgcatgccgc
     5461 ttccccgact cggtgccact ttttcgggag gcagataggt aggcatggcc cccattttca
     5521 atacaagcaa cgcatgagaa agcccccgga agatcacctt ccgggggctt ttttattgcg
     5581 ctccggcaat taaaaaagcg gctaaccacg ccgctttttt tacgtctgca gactagtcag
     5641 gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa agacgtctca ttaatcagat
     5701 aaaatattga caatgtgagc ggataacaag atactgagca catcagcagc acacaggaaa
     5761 cagaccatgg aattcgagct cggtacccgg ggatcctcta gagtcgacct gcaggcatgc
     5821 aagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa
     5881 cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct
     5941 gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc
     6001 catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg
     6061 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
     6121 gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc
     6181 ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc ttttcacaga
     6241 atgagcatca tggcctagtc gacatcggag aagagtacgg ctcttttaac cgcctcaaga
     6301 accagataag tgaaatctag ttccaaacta ttttgtcatt tttaattttc gtattagctt
     6361 acgacgctac acccagttcc catctatttt gtcactcttc cctaaataat ccttaaaaac
     6421 tccatttcca cccctcccag ttcccaacta ttttgtccgc ccacagcggg gcatttttct
     6481 tcctgttatg tttgggctct tccgcttcct cgctcactga ccctatcaga gcttatcggc
     6541 cagcctcgca gagcaggatt cccgttgagc accgccaggt gcgaataagg gacagtgaag
     6601 aaggaacacc cgctcgcggg tgggcctact tcacctatcc tgcccggctg acgccgttgg
     6661 atacaccaag gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag
     6721 gatggatata ccgaaaaaat cgctataatg accccgaagc agggttatgc agcggaaagt
     6781 ataccttaag gaatcccctg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6841 ggccaggaac cgtaaaaagg ccgcactccc tgcctctgtc atcacgatac tgtgatgccg
     6901 tggctaattc ccatgtcagc cgttaagtgt tcctgtgtca ctcaaaattg ctttgagagg
     6961 ctctaagggc ttctcagtgc gttacatccc tggcttgttg tccacaaccg ttaaacctta
     7021 aaagctttaa aagccttata tattcttttt tttcttataa aacttaaaac cttagaggct
     7081 atttaagttg ctgatttata ttaattttat tgttcaaaca tgagagctta gtacgtgaaa
     7141 catgagagct tagtacgtta gccatgagag cttagtacgt tagccatgag ggtttagttc
     7201 gttaaacatg agagcttagt acgttaaaca tgagagctta gtacgtgaaa catgagagct
     7261 tagtacgtac tatcaacagg ttgaactgct gatcttcaga tcggcc
//
