LOCUS       sequence-161171-        7261 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mini-Tn7 delivery plasmid with metE under the cpcG2-Delta-59 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7261)
  AUTHORS   Milias-Argeitis A, Rullan M, Aoki SK, Buchmann P, Khammash M
  TITLE     Automated optogenetic feedback control for precise and robust 
            regulation of gene expression and cell growth.
  JOURNAL   Nat Commun. 2016 Aug 26;7:12546. doi: 10.1038/ncomms12546.
  PUBMED    27562138
REFERENCE   2  (bases 1 to 7261)
  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: "Nat 
            Commun."; date: "2016-08-26"; volume: "7:12546. doi"; pages: " 
            10.1038/ncomms12546"
COMMENT     Sequence Label: pSKA397
FEATURES             Location/Qualifiers
     source          1..7261
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(114..161)
                     /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(178..837)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     752..771
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(838..940)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(965..998)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(1432..1451)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1551..1573
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1611..1629)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1697..1801
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1802..1870,1871..2662)
                     /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     complement(2020..2039)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3071..3427
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            3478..3586
                     /note="incP origin of transfer"
     mobile_element  3890..4055
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     terminator      complement(4118..4145)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4237..4323)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4476..4493
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     RBS             6806..6817
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      complement(7048..7134)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(join(7237..7261,1..70))
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc
       61 aacaggagtc caagctcagc taattaagct agcgtgtagg ctggagctgc ttcgaagttc
      121 ctatactttc tagagaatag gaacttcgga ataggaactt catttaaatg gcgcgcctta
      181 cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg
      241 gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc
      301 ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac
      361 gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc
      421 aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata
      481 tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc
      541 agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc
      601 gtctttcatt gccatacgta attccggatg agcattcatc aggcgggcaa gaatgtgaat
      661 aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc
      721 cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc
      781 tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt
      841 agcttcctta gctcctgaaa atctcgacaa ctcaaaaaat acgcccggta gtgatcttat
      901 ttcattatgg tgaaagttgg aacctcttac gtgccgatca acgtctcatt ttcgccaaaa
      961 gttggaagtt cctatacttt ctagagaata ggaacttcga actgcaggtc gacctcgaac
     1021 cccacgcccc tctttaatac gacgggcaat ttgcacttca gaaaatgaag agtttgcttt
     1081 agccataaca aaagtccagt atgctttttc acagcataac tggactgatt tcagtttaca
     1141 actattctgt ctagtttaag actttattgt catagtttag atctattttg ttcagtttaa
     1201 gactttattg tccgcctaca cccgcttacg cagggcatcc atttattact caaccgtaac
     1261 cgattttgcc aggttacgcg gctggtcctc tagctggcgt aatagcgaag aggcccgcac
     1321 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt
     1381 tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg
     1441 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     1501 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     1561 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     1621 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     1681 ttttcgggga aatgtacgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     1741 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     1801 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     1861 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     1921 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     1981 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     2041 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     2101 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     2161 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     2221 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     2281 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     2341 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     2401 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     2461 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     2521 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     2581 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     2641 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     2701 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     2761 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2821 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2881 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2941 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3001 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3061 accagccggt tgtcagccgt taagtgttcc tgtgtcactc aaaattgctt tgagaggctc
     3121 taagggcttc tcagtgcgtt acatccctgg cttgttgtcc acaaccgtta aaccttaaaa
     3181 gctttaaaag ccttatatat tctttttttt cttataaaac ttaaaacctt agaggctatt
     3241 taagttgctg atttatatta attttattgt tcaaacatga gagcttagta cgtgaaacat
     3301 gagagcttag tacgttagcc atgagagctt agtacgttag ccatgagggt ttagttcgtt
     3361 aaacatgaga gcttagtacg ttaaacttga gagcttagta cgtgaaacat gagagcttag
     3421 tacgtactat caacaggttg aactgcccat gttctttcct gcgttatcag agcttatcgg
     3481 ccagcctcgc agagcaggat tcccgttgag caccgccagg tgcgaataag ggacagtgaa
     3541 gaaggaacac ccgctcgcgg gtgggcctac ttcacctatc ctgcccggct gacgccgttg
     3601 gatacaccaa ggaaagtcta cacgaaccct ttggcaaaat cctgtatatc gtgcgaaaaa
     3661 ggatggatat accgaaaaaa tcgctataat gaccccgaag cagggttatg cagcggaaag
     3721 tataccttaa ggaatcccct gataccgctc gccgcagccg aacgaccgag cgcagcgggt
     3781 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     3841 cgattcatta atgcagcgcc caaacataac aggaagaaaa atgccccgct gtgggcggac
     3901 aaaatagttg ggaactggga ggggtggaaa tggagttttt aaggattatt tagggaagag
     3961 tgacaaaata gatgggaact gggtgtagcg tcgtaagcta atacgaaaat taaaaatgac
     4021 aaaatagttt ggaactagat ttcacttatc tggttggcct gcaaggcctt cgcgaggtac
     4081 cgggcccaag cttctcgaga gagtttgtag aaacgcaaaa aggccatccg tcaggatggc
     4141 cttctgctta atttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg
     4201 ccgttgcttc gcaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt
     4261 caccgacaaa caacagataa aacgaaaggc ccagtctttc gactgagcct ttcgttttat
     4321 ttgatgcctg gcagttccct actctcgcat ggggagaccc cacactacca tcggcgctac
     4381 ggcgtttcac ttctgagttc ggcatggggt caggtgggac caccgcgcta ctgccgccag
     4441 gcaaattctg ttttatcaga ccgcttctgc gttctgattt aatctgtatc aggctgaaaa
     4501 tcttctctca tccgccaaaa cagccaagct tgcatgctta cccccgacgc aagttctgcg
     4561 ccgcctgcac catgttcgcc agtgccgcgc gggtttctgg ccagccgcgc gttttcaggc
     4621 cacagtccgg gttgacccac aggcgctctg ccggaatgcg ttttgccgct ttcttcagca
     4681 aggcttcaat ccattccacg ctcggtacgt ttggcgagtg aatgtcatag acgccaggac
     4741 cgatttcatt tggataatca aactcttcaa acgactccag caactccatg tcggaacgcg
     4801 aggtttcgat ggtgatgacg tctgcgtcca gcgccgcaat cgaatccatg atgtcgttga
     4861 actcgcaata acacatgtga gtgtggattt gtgtgtcatc tttcgccacg gcggcgttga
     4921 tacggaaggc ctctacgccc cactggagat acgcatccca gtcgctacga cgcagcggta
     4981 aaccttcgcg cagcgccggt tcgtcaatct ggatgatgcc aattccagcg gcttccagat
     5041 cggccacttc atcacgcagc gccagcgcaa tctgtttggc gatggtttca cggctgacat
     5101 cttcacgcgg gaacgaccag cagagtatgg tcaccggccc cgtcagcatc cctttcaccg
     5161 gtttgtcggt cagcgattgc gcatacttcg cccactccac ggtaatcggt gccgggcggc
     5221 taatgtcacc aatgacaatc ggtggcttca cgcagcggga accgtagctc tgtacccaac
     5281 cgttttgcgt aaagacaaat ccgtcgaggt gctcgccaaa gtattccacc atgtcattac
     5341 gctcggcctc gccatgtacc agcacatcca gtcccaaacg ttcctgctca acaatggcct
     5401 gcttgatatg ttccgcaatg cccgtgcggt agttgttggc gtcgagattg ccctttttga
     5461 aatccagacg cagggtacga atttccgtgg tttgcgggaa ggaaccaatc gtggtggtcg
     5521 gccacgctgg cagtttaaaa cgcgcacgct gggcttcagc acgcacttca tagacattcg
     5581 cacgctggct gtcctgggcg gtgatcgccg ccagacgctt ttctaccgcc ggattatgta
     5641 cgcgggtaga gtgacgacgt gcctgaatcg gggcgctcca ctctgccaga gctgccgtgt
     5701 caccactgtt cagcgcatcg cgcagcagtg ccagttcatg gcatttttgt agggcgaagg
     5761 caaaccagct tttcacttct gcatcaagac gcgtttccac gctcaggtcg atggggctgt
     5821 gcagcaacga gcaggaagat gccacccaca aatcacgttt gccgacaatg tccttaattt
     5881 gcgcatattt ctcggtaaga tcggcgcgcc agacgttacg accattgatc agacccgcag
     5941 acagcaacca gtcagaaggc aggcgcttgt gcagttcagc aacgtcatct ttaccatgta
     6001 cgaggtcaac atgcagaccc tgaacaggca gcgcagtaat cgtgtcgaga tttggcgtta
     6061 cgccttcaaa ataggtggtc agcagcagtt tcacctgtcc ctggagcgcg tcgtaagcgg
     6121 gtttgtatgc gtccagccac gcctgtggta gttccagtac cagcgcgggt tcatcaatct
     6181 gtacccactc gatgccgcgt ttcgccagtt ctgccagcac ttgctgataa accggcagaa
     6241 tgtcgttcag caggctcagg cggtcaaatt gttcaccttt cactttcccc agccacagcc
     6301 aggtaaccgg ccccagcagc acaggtttca ccttgtggcc cagcgccagc gcctcgtcca
     6361 cttcgtccag cagctgcgtc caggtcagtt tgaactgttg gcctttaacg aactccggca
     6421 ccatgtagtg atagttggtg ttaaaccatt tggtcatttc cgctgccgcc gcaggttcgc
     6481 cagtcggcgc acgtccacga ccaatacgga acagggtgtc gatatctacc gaaccatctt
     6541 tgttctgatg acgcgccgga acgttaccca gcagcagact ggtggtcagt acatgatcgt
     6601 accaggcaaa atcgcccacc ggcagcaggt cgatacccgc ttgcttttgt tgatcccagt
     6661 gacgagcacg caattcacgc cctaccgcca gcagttcttc acgcgtggag ttccccgccc
     6721 aataactttc ttgcgctttt ttcagctcgc gacgcaggcc aacgcgaggg aaaccgaggg
     6781 tgtgattcaa tattgtcatc tagtatttct cctctttaga taaagttaga atgcgatcct
     6841 aacaaagtaa aattgaagaa aagggggaga aaatcgtaaa gaaatcggaa agttagctga
     6901 tgcgactaaa actgttacat ttcgcaacat gaattaaccc agacctgttc accccttcag
     6961 gtaagctgag gttgatagag aaaagcacaa tgggctacta gtgagctcat gcatgatatc
     7021 gtccgggccg caagctccta gcggcggatt tgtcctactc aggagagcgt tcaccgacaa
     7081 acaacagata aaacgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgcct
     7141 caagctagag agtcattacc ccaggcgttt aagggcacca ataactgcct taaaaaaatt
     7201 acgccccgcc ctgccactca tcgcagtcta gcttggattc tcaccaataa aaaacgcccg
     7261 g
//
