LOCUS       sequence-222797-        6076 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Periplasmic expression of superfolder mTurquoise2ox (pNM077 
            pTHV037-DsbAss-LEGPAGL-sfTq2ox-EFGS-∆1-17-PBP5).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6076)
  AUTHORS   Meiresonne NY, Consoli E, Mertens LMY, Chertkova AO, Goedhart J, den
            Blaauwen T
  TITLE     Superfolder mTurquoise2(ox) optimized for the bacterial periplasm 
            allows high efficiency in vivo FRET of cell division antibiotic 
            targets.
  JOURNAL   Mol Microbiol. 2019 Apr;111(4):1025-1038. doi: 10.1111/mmi.14206. 
            Epub 2019 Feb 28.
  PUBMED    30648295
REFERENCE   2  (bases 1 to 6076)
  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"; doi: "10.1111/mmi.14206";
            journalName: "Mol Microbiol"; date: "2019-04"; volume: "111"; issue:
            "4"; pages: "1025-1038"
COMMENT     Sequence Label: sfTq2ox-PBP5
FEATURES             Location/Qualifiers
     source          1..6076
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(272..277,278..294,295..301)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    309..325
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by 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)."
     primer_bind     314..336
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             join(442..444,445..447,448..1158)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQVFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNVEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(487..508)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1095..1116
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2338..2355)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2508..2594
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2686..2713
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2733..2824
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2825..2893,2894..3685)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3043..3062)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3856..4444
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4345..4364
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4598..4615
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4630..4770
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4696..4715)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4856..4878
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4956..5033
                     /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."
     CDS             join(5034..6076,1..40)
                     /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"
     primer_bind     complement(5053..5072)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
       61 gttagcgcga attgatctgg tttgacagct tatcatcgac tgcacggtgc accaatgctt
      121 ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcactgcat
      181 aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata
      241 acggttctgg caaatattct gaaatgagct gtttacaatt aatcatccgg ctcgtataat
      301 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agaccatggg caaaaagatt
      361 tggctggcgc tggctggttt agttttagcg tttagcgcat cggcggcgca gtatgaagat
      421 ctcgagggtc cggctggtct gatggtgagc aagggcgagg agctgttcac cggggtggtg
      481 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gcgcggcgag
      541 ggcgagggcg atgccaccaa tggcaagctg accctgaagt tcatctgcac caccggcaag
      601 ctgcccgtgc cctggcccac cctcgtgacc accctgtcct ggggtgtaca ggtcttcgcc
      661 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
      721 gtccaggagc gcaccatcag cttcaaggac gacggcacat acaagacccg cgccgaggtg
      781 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
      841 gacggcaaca tcctggggca caagctggag tacaactact ttagcgacaa cgtctatatc
      901 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacgtggag
      961 gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1021 gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac
     1081 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1141 atggacgagc tgtacaagga attcggatcc gatgacctga atatcaaaac tatgatcccg
     1201 ggtgtaccgc agatcgatgc ggagtcctac atcctgattg actataactc cggcaaagtg
     1261 ctcgccgaac agaacgcaga tgtccgccgc gatcctgcca gcctgaccaa aatgatgacc
     1321 agttacgtta tcggccaggc aatgaaagcc ggtaaattta aagaaactga tttagtcact
     1381 atcggcaacg acgcatgggc caccggtaac ccggtgttta aaggttcttc gctgatgttc
     1441 ctcaaaccgg gcatgcaggt tccggtttct cagctgatcc gcggtattaa cctgcaatcg
     1501 ggtaacgatg cttgtgtcgc catggccgat tttgccgctg gtagccagga cgcttttgtt
     1561 ggcttgatga acagctacgt taacgcactg ggcctgaaaa atacccactt ccagacggta
     1621 catggtctgg atgctgatgg tcagtacagc tccgcgcgag atatggcgct gatcggccag
     1681 gcattgatcc gtgacgtacc gaatgaatac tcgatctata aagaaaaaga atttacgttt
     1741 aacggtattc gccagctgaa ccgtaacggc ctgttatggg ataacagcct gaatgtcgac
     1801 ggcatcaaaa ccggacacac tgacaaagca ggttacaacc ttgttgcttc tgcgactgaa
     1861 ggccagatgc gcttgatttc tgcggtaatg ggcggacgta cttttaaagg ccgtgaagcc
     1921 gaaagtaaaa aactgctaac ctggggcttc cgtttctttg aaaccgttaa cccactgaaa
     1981 gtaggtaaag agttcgcctc tgaaccggtt tggtttggtg attctgatcg cgcttcgtta
     2041 ggggttgata aagacgtgta cctgaccatt ccgcgtggtc gcatgaaaga tctgaaagcc
     2101 agctatgtgc tgaacagcag tgaattgcat gcgccgctgc aaaagaatca ggtcgtcgga
     2161 actatcaact tccagcttga tggcaaaacg atcgagcaac gcccgctggt tgtgttgcaa
     2221 gaaatcccgg aaggtaactt cttcggcaaa atcattgatt acattaaatt aatgttccat
     2281 cactggtttg gttaaaaaga agcttggctg ttttggcgga tgagagaaga ttttcagcct
     2341 gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag
     2401 tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga
     2461 tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa
     2521 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     2581 tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt
     2641 ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga
     2701 cggatggcct ttttgcgttt ctacaaactc tttttgttta tttttctaaa tacattcaaa
     2761 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2821 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     2881 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     2941 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     3001 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     3061 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     3121 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     3181 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     3241 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     3301 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3361 gatgcctaca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3421 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3481 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     3541 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     3601 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3661 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     3721 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3781 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3841 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3901 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3961 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     4021 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4081 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4141 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4201 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4261 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4321 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4381 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4441 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4501 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     4561 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     4621 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     4681 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
     4741 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     4801 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     4861 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gcagatcaat
     4921 tcgcgcgcga aggcgaagcg gcatgcattt acgttgacac catcgaatgg tgcaaaacct
     4981 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac
     5041 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg
     5101 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg
     5161 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc
     5221 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga
     5281 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg
     5341 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga
     5401 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg
     5461 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc
     5521 atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg
     5581 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata
     5641 aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca
     5701 tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc
     5761 tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc
     5821 gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata
     5881 tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc
     5941 gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac
     6001 tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg
     6061 ccgattcatt aatgca
//
