LOCUS       sequence-151229-        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"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    604..637
                     /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)"
     promoter        662..764
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             765..1424
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(831..850)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    1441..1488
                     /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      1532..1626
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      1729..1815
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             2046..2057
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(4370..4387)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4540..4626
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4718..4745
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     mobile_element  4808..4973
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            5277..5385
                     /note="incP origin of transfer"
     rep_origin      5436..5792
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(join(6201..6992,6993..7061))
                     /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     6824..6843
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7062..7166)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7234..7252
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct agaggaccag ccgcgtaacc tggcaaaatc ggttacggtt gagtaataaa
      361 tggatgccct gcgtaagcgg gtgtaggcgg acaataaagt cttaaactga acaaaataga
      421 tctaaactat gacaataaag tcttaaacta gacagaatag ttgtaaactg aaatcagtcc
      481 agttatgctg tgaaaaagca tactggactt ttgttatggc taaagcaaac tcttcatttt
      541 ctgaagtgca aattgcccgt cgtattaaag aggggcgtgg ggttcgaggt cgacctgcag
      601 ttcgaagttc ctattctcta gaaagtatag gaacttccaa cttttggcga aaatgagacg
      661 ttgatcggca cgtaagaggt tccaactttc accataatga aataagatca ctaccgggcg
      721 tattttttga gttgtcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca
      781 ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc
      841 agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa
      901 agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc
      961 tgatgaatgc tcatccggaa ttacgtatgg caatgaaaga cggtgagctg gtgatatggg
     1021 atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct
     1081 ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt
     1141 gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct
     1201 cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact
     1261 tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc
     1321 cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta
     1381 atgaattaca acagtactgc gatgagtggc agggcggggc gtaaggcgcg ccatttaaat
     1441 gaagttccta ttccgaagtt cctattctct agaaagtata ggaacttcga agcagctcca
     1501 gcctacacgc tagcttaatt agctgagctt ggactcctgt tgatagatcc agtaatgacc
     1561 tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt
     1621 gagaatccaa gctagactgc gatgagtggc agggcggggc gtaatttttt taaggcagtt
     1681 attggtgccc ttaaacgcct ggggtaatga ctctctagct tgaggcatca aataaaacga
     1741 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     1801 ctgagtagga caaatccgcc gctaggagct tgcggcccgg acgatatcat gcatgagctc
     1861 actagtagcc cattgtgctt ttctctatca acctcagctt acctgaaggg gtgaacaggt
     1921 ctgggttaat tcatgttgcg aaatgtaaca gttttagtcg catcagctaa ctttccgatt
     1981 tctttacgat tttctccccc ttttcttcaa ttttactttg ttaggatcgc attctaactt
     2041 tatctaaaga ggagaaatac tagatgacaa tattgaatca caccctcggt ttccctcgcg
     2101 ttggcctgcg tcgcgagctg aaaaaagcgc aagaaagtta ttgggcgggg aactccacgc
     2161 gtgaagaact gctggcggta gggcgtgaat tgcgtgctcg tcactgggat caacaaaagc
     2221 aagcgggtat cgacctgctg ccggtgggcg attttgcctg gtacgatcat gtactgacca
     2281 ccagtctgct gctgggtaac gttccggcgc gtcatcagaa caaagatggt tcggtagata
     2341 tcgacaccct gttccgtatt ggtcgtggac gtgcgccgac tggcgaacct gcggcggcag
     2401 cggaaatgac caaatggttt aacaccaact atcactacat ggtgccggag ttcgttaaag
     2461 gccaacagtt caaactgacc tggacgcagc tgctggacga agtggacgag gcgctggcgc
     2521 tgggccacaa ggtgaaacct gtgctgctgg ggccggttac ctggctgtgg ctggggaaag
     2581 tgaaaggtga acaatttgac cgcctgagcc tgctgaacga cattctgccg gtttatcagc
     2641 aagtgctggc agaactggcg aaacgcggca tcgagtgggt acagattgat gaacccgcgc
     2701 tggtactgga actaccacag gcgtggctgg acgcatacaa acccgcttac gacgcgctcc
     2761 agggacaggt gaaactgctg ctgaccacct attttgaagg cgtaacgcca aatctcgaca
     2821 cgattactgc gctgcctgtt cagggtctgc atgttgacct cgtacatggt aaagatgacg
     2881 ttgctgaact gcacaagcgc ctgccttctg actggttgct gtctgcgggt ctgatcaatg
     2941 gtcgtaacgt ctggcgcgcc gatcttaccg agaaatatgc gcaaattaag gacattgtcg
     3001 gcaaacgtga tttgtgggtg gcatcttcct gctcgttgct gcacagcccc atcgacctga
     3061 gcgtggaaac gcgtcttgat gcagaagtga aaagctggtt tgccttcgcc ctacaaaaat
     3121 gccatgaact ggcactgctg cgcgatgcgc tgaacagtgg tgacacggca gctctggcag
     3181 agtggagcgc cccgattcag gcacgtcgtc actctacccg cgtacataat ccggcggtag
     3241 aaaagcgtct ggcggcgatc accgcccagg acagccagcg tgcgaatgtc tatgaagtgc
     3301 gtgctgaagc ccagcgtgcg cgttttaaac tgccagcgtg gccgaccacc acgattggtt
     3361 ccttcccgca aaccacggaa attcgtaccc tgcgtctgga tttcaaaaag ggcaatctcg
     3421 acgccaacaa ctaccgcacg ggcattgcgg aacatatcaa gcaggccatt gttgagcagg
     3481 aacgtttggg actggatgtg ctggtacatg gcgaggccga gcgtaatgac atggtggaat
     3541 actttggcga gcacctcgac ggatttgtct ttacgcaaaa cggttgggta cagagctacg
     3601 gttcccgctg cgtgaagcca ccgattgtca ttggtgacat tagccgcccg gcaccgatta
     3661 ccgtggagtg ggcgaagtat gcgcaatcgc tgaccgacaa accggtgaaa gggatgctga
     3721 cggggccggt gaccatactc tgctggtcgt tcccgcgtga agatgtcagc cgtgaaacca
     3781 tcgccaaaca gattgcgctg gcgctgcgtg atgaagtggc cgatctggaa gccgctggaa
     3841 ttggcatcat ccagattgac gaaccggcgc tgcgcgaagg tttaccgctg cgtcgtagcg
     3901 actgggatgc gtatctccag tggggcgtag aggccttccg tatcaacgcc gccgtggcga
     3961 aagatgacac acaaatccac actcacatgt gttattgcga gttcaacgac atcatggatt
     4021 cgattgcggc gctggacgca gacgtcatca ccatcgaaac ctcgcgttcc gacatggagt
     4081 tgctggagtc gtttgaagag tttgattatc caaatgaaat cggtcctggc gtctatgaca
     4141 ttcactcgcc aaacgtaccg agcgtggaat ggattgaagc cttgctgaag aaagcggcaa
     4201 aacgcattcc ggcagagcgc ctgtgggtca acccggactg tggcctgaaa acgcgcggct
     4261 ggccagaaac ccgcgcggca ctggcgaaca tggtgcaggc ggcgcagaac ttgcgtcggg
     4321 ggtaagcatg caagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga
     4381 ttaaatcaga acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg
     4441 tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg
     4501 tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag
     4561 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     4621 acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca
     4681 ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc
     4741 ctttttgcgt ttctacaaac tctctcgaga agcttgggcc cggtacctcg cgaaggcctt
     4801 gcaggccaac cagataagtg aaatctagtt ccaaactatt ttgtcatttt taattttcgt
     4861 attagcttac gacgctacac ccagttccca tctattttgt cactcttccc taaataatcc
     4921 ttaaaaactc catttccacc cctcccagtt cccaactatt ttgtccgccc acagcggggc
     4981 atttttcttc ctgttatgtt tgggcgctgc attaatgaat cggccaacgc gcggggagag
     5041 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgacccgctg cgctcggtcg
     5101 ttcggctgcg gcgagcggta tcaggggatt ccttaaggta tactttccgc tgcataaccc
     5161 tgcttcgggg tcattatagc gattttttcg gtatatccat cctttttcgc acgatataca
     5221 ggattttgcc aaagggttcg tgtagacttt ccttggtgta tccaacggcg tcagccgggc
     5281 aggataggtg aagtaggccc acccgcgagc gggtgttcct tcttcactgt cccttattcg
     5341 cacctggcgg tgctcaacgg gaatcctgct ctgcgaggct ggccgataag ctctgataac
     5401 gcaggaaaga acatgggcag ttcaacctgt tgatagtacg tactaagctc tcatgtttca
     5461 cgtactaagc tctcaagttt aacgtactaa gctctcatgt ttaacgaact aaaccctcat
     5521 ggctaacgta ctaagctctc atggctaacg tactaagctc tcatgtttca cgtactaagc
     5581 tctcatgttt gaacaataaa attaatataa atcagcaact taaatagcct ctaaggtttt
     5641 aagttttata agaaaaaaaa gaatatataa ggcttttaaa gcttttaagg tttaacggtt
     5701 gtggacaaca agccagggat gtaacgcact gagaagccct tagagcctct caaagcaatt
     5761 ttgagtgaca caggaacact taacggctga caaccggctg gtaacaggat tagcagagcg
     5821 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5881 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5941 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6001 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6061 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6121 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6181 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6241 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6301 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6361 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6421 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6481 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6541 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6601 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6661 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6721 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6781 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6841 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6901 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6961 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7021 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7081 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7141 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     7201 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     7261 c
//
