LOCUS       pJMP12042        7108 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 7108)
  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..7108
                     /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..6267
                     /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..3185)
                     /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"
     promoter        complement(join(3374..3379,3380..3396,3397..3402))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    3519..3566
                     /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(3567..3656)
                     /label=spy terminator
     terminator      complement(3745..3795)
                     /label=L3S1P13 terminator
     terminator      complement(3755..3795)
                     /label=L3S1P13 terminator
     terminator      complement(3882..3938)
                     /label=thrLABC terminator
     terminator      complement(3939..3992)
                     /label=garPLRK-rnpB terminator
     CDS             complement(4084..5166)
                     /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        5167..5244
                     /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      5331..5383
                     /label=hisLGDCBHAFI terminator
     terminator      5384..5432
                     /label=pdhR-aceEF-lpd terminator
     misc_feature    join(5509..5515,5516..5531,5532..5537)
                     /label=PabstBR promoter
     misc_feature    5516..5531
                     /label=lac operator
     misc_feature    5539..5568
                     /label=Gibson ends - NcoI/BamHI cloning
     misc_feature    5572..5628
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     misc_feature    5594..5623
                     /label=Gibson ends - NcoI/BamHI cloning
     terminator      5711..6036
                     /label=rrnB terminators
                     /label=rrnB
     terminator      5831..5917
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6009..6036
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     mobile_element  6102..6267
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     misc_feature    6326..6583
                     /label=expanded oriT
     oriT            6339..6447
                     /note="incP origin of transfer"
     rep_origin      6716..7104
                     /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 gttagccaat cgactggcga
     2401 gcggcatcgc attcttcgca tcccgcctct ggcggatgca ggaagatcaa cggatctcgg
     2461 cccagttgac ccagggctgt cgccacaatg tcgcgggagc ggatcaaccg agcaaaggca
     2521 tgaccgactg gaccttcctt ctgaaggctg ttctccttga gccacctgtc cgccaaggca
     2581 aagcgctcac agcagtggtc attctccaga taatcgacac gtaccaactt gccatcctga
     2641 agaatggtgc agtgtctcgg caccccatag ggaacctttg ccatcaactc ggcaagatgc
     2701 agcgtcgtgt tggcatcgtg tcccacgccg aggagaagta cctgcccatc gagttcatgg
     2761 acacgggcga ccgggcttgc aggcgagtga ggtggcaggg gcaatggatc agagatgatc
     2821 tgctctgcct gtggccccgc tgccgcaaag gcaaatggat gggcgctgcg ctttacattt
     2881 ggcagacgcc agaatgtgtc agagacaact ccaaggtccg gtgtaacggg cgacgtggca
     2941 ggatcgaacg gctcgtcgtc cagacctgac cacgatggca tgacgagcgt ccctcccgga
     3001 cccagtgcag cacgcagggc ctcgatcagt ccaagtggcc catcttcgag gggccggacg
     3061 ctacggaagg agctgtggac cagcagcaca ccgccggggg taaccccaag gttgagaagc
     3121 tgaccgatca gctcggcttt tcgccattcg tattgcacga cattgcactc caccgctgat
     3181 gacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt
     3241 ggatgcccga ggcatagact gtacgaaaaa acagtcataa caagccatga aaaccgccac
     3301 tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg agcgcatacg
     3361 ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttc gtgccttcat
     3421 ccgtttccac ggtgtgcgtc catgagcaaa tattatacgc aaggcgacaa ggctcgagag
     3481 ttcaacagac agccttcgtt cttatggccg ttcaggagga agttcctatt ccgaagttcc
     3541 tattctctag aaagtatagg aacttcttga gaagagaaaa gaaaaccgcc gatcctgtcc
     3601 accgcattac tgcaaggtag tggacaagac cggcggtctt aagttttttg gctgaaacga
     3661 atgacgaaaa ggctgtctga gcaaatccag gaggtcgttt aaaccgtatt agcccggtat
     3721 tttggaaata gcggaagcac tgacttttgt tatcaataaa aaaggccccc cgttagggag
     3781 gccttattgt tcgtctactc ggaagagcga gagacaacag aacggtcagc cacatgaatt
     3841 ctttcagctc agtcgatagg tagtaggcaa gagtagtcgc acctttggtc gaaaaaaaaa
     3901 gcccgcactg tcaggtgcgg gcttttttct gtgtttcccc aaaagtaaaa acccgccgaa
     3961 gcgggttttt acgtaaaaca ggtgaaactg accagacgag aaggctttgg aggtgataat
     4021 ggggctcaag gacccgggtg gggtgcctaa tgagtgagct aacttacatt aattgcgttg
     4081 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4141 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     4201 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     4261 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     4321 ataacatgag ctatcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg
     4381 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     4441 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     4501 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     4561 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     4621 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac catcttcatg
     4681 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     4741 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     4801 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     4861 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     4921 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     4981 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     5041 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     5101 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     5161 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     5221 ggttttgcac cattcgatgg tgtcaacgta aatgcatgcc gcttccccga ctcggtgcca
     5281 ctttttcggg aggcagatag gtaggcatgg cccccatttt caatacaagc aacgcatgag
     5341 aaagcccccg gaagatcacc ttccgggggc ttttttattg cgctccggca attaaaaaag
     5401 cggctaacca cgccgctttt tttacgtctg cagactagtc aggcagccat cggaagctgt
     5461 ggtatggctg tgcaggtcgt aaagacgtct cattaatcag ataaaatatt gacaatgtga
     5521 gcggataaca agatactgag cacatcagca gcacacagga aacagaccat ggaattcgag
     5581 ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat gcaagcttgg ctgttttggc
     5641 ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata
     5701 aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca
     5761 gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac
     5821 tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     5881 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt
     5941 tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca
     6001 aattaagcag aaggccatcc tgacggatgg ccttttcaca gaatgagcat catggcctag
     6061 tcgacatcgg agaagagtac ggctctttta accgcctcaa gaaccagata agtgaaatct
     6121 agttccaaac tattttgtca tttttaattt tcgtattagc ttacgacgct acacccagtt
     6181 cccatctatt ttgtcactct tccctaaata atccttaaaa actccatttc cacccctccc
     6241 agttcccaac tattttgtcc gcccacagcg gggcattttt cttcctgtta tgtttgggct
     6301 cttccgcttc ctcgctcact gaccctatca gagcttatcg gccagcctcg cagagcagga
     6361 ttcccgttga gcaccgccag gtgcgaataa gggacagtga agaaggaaca cccgctcgcg
     6421 ggtgggccta cttcacctat cctgcccggc tgacgccgtt ggatacacca aggaaagtct
     6481 acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa
     6541 atcgctataa tgaccccgaa gcagggttat gcagcggaaa gtatacctta aggaatcccc
     6601 tgataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6661 ggccgcactc cctgcctctg tcatcacgat actgtgatgc cgtggctaat tcccatgtca
     6721 gccgttaagt gttcctgtgt cactcaaaat tgctttgaga ggctctaagg gcttctcagt
     6781 gcgttacatc cctggcttgt tgtccacaac cgttaaacct taaaagcttt aaaagcctta
     6841 tatattcttt tttttcttat aaaacttaaa accttagagg ctatttaagt tgctgattta
     6901 tattaatttt attgttcaaa catgagagct tagtacgtga aacatgagag cttagtacgt
     6961 tagccatgag agcttagtac gttagccatg agggtttagt tcgttaaaca tgagagctta
     7021 gtacgttaaa catgagagct tagtacgtga aacatgagag cttagtacgt actatcaaca
     7081 ggttgaactg ctgatcttca gatcggcc
//
