LOCUS       sequence-227549-        6075 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Periplasmic expression of superfolder mTurquoise2 (pNM067 
            pTHV037-DsbAss-LEGPAGL-sfTq2-EFGS-∆1-17-PBP5).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6075)
  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 6075)
  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: sfTq2-PBP5
FEATURES             Location/Qualifiers
     source          1..6075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(102..107,108..124,125..131)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    139..155
                     /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     144..166
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             join(272..274,275..277,278..988)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MGSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNVEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(317..338)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     925..946
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2168..2185)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2338..2424
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2516..2543
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2562..2653
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2654..2722,2723..3514)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2872..2891)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3685..4273
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4174..4193
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4427..4444
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4459..4599
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4525..4544)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4685..4707
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4785..4862
                     /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             4863..5945
                     /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(4882..4901)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc
       61 cgacatcata acggttctgg caaatattct gaaatgagct gtttacaatt aatcatccgg
      121 ctcgtataat gtgtggaatt gtgagcggat aacaatttca cacaggaaac agaccatggg
      181 caaaaagatt tggctggcgc tggctggttt agttttagcg tttagcgcat cggcggcgca
      241 gtatgaagat ctcgagggtc cggctggtct gatggggagc aagggcgagg agctgttcac
      301 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
      361 gcgcggcgag ggcgagggcg atgccaccaa tggcaagctg accctgaagt tcatctgcac
      421 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgtcct ggggcgtgca
      481 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
      541 cgaaggctac gtccaggagc gcaccatcag cttcaaggac gacggcacat acaagacccg
      601 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
      661 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactact ttagcgacaa
      721 cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca
      781 caacgtggag gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
      841 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa
      901 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
      961 cactctcggc atggacgagc tgtacaagga attcggatcc gatgacctga atatcaaaac
     1021 tatgatcccg ggtgtaccgc agatcgatgc ggagtcctac atcctgattg actataactc
     1081 cggcaaagtg ctcgccgaac agaacgcaga tgtccgccgc gatcctgcca gcctgaccaa
     1141 aatgatgacc agttacgtta tcggccaggc aatgaaagcc ggtaaattta aagaaactga
     1201 tttagtcact atcggcaacg acgcatgggc caccggtaac ccggtgttta aaggttcttc
     1261 gctgatgttc ctcaaaccgg gcatgcaggt tccggtttct cagctgatcc gcggtattaa
     1321 cctgcaatcg ggtaacgatg cttgtgtcgc catggccgat tttgccgctg gtagccagga
     1381 cgcttttgtt ggcttgatga acagctacgt taacgcactg ggcctgaaaa atacccactt
     1441 ccagacggta catggtctgg atgctgatgg tcagtacagc tccgcgcgag atatggcgct
     1501 gatcggccag gcattgatcc gtgacgtacc gaatgaatac tcgatctata aagaaaaaga
     1561 atttacgttt aacggtattc gccagctgaa ccgtaacggc ctgttatggg ataacagcct
     1621 gaatgtcgac ggcatcaaaa ccggacacac tgacaaagca ggttacaacc ttgttgcttc
     1681 tgcgactgaa ggccagatgc gcttgatttc tgcggtaatg ggcggacgta cttttaaagg
     1741 ccgtgaagcc gaaagtaaaa aactgctaac ctggggcttc cgtttctttg aaaccgttaa
     1801 cccactgaaa gtaggtaaag agttcgcctc tgaaccggtt tggtttggtg attctgatcg
     1861 cgcttcgtta ggggttgata aagacgtgta cctgaccatt ccgcgtggtc gcatgaaaga
     1921 tctgaaagcc agctatgtgc tgaacagcag tgaattgcat gcgccgctgc aaaagaatca
     1981 ggtcgtcgga actatcaact tccagcttga tggcaaaacg atcgagcaac gcccgctggt
     2041 tgtgttgcaa gaaatcccgg aaggtaactt cttcggcaaa atcattgatt acattaaatt
     2101 aatgttccat cactggtttg gttaaaaaga agcttggctg ttttggcgga tgagagaaga
     2161 ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc
     2221 ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc
     2281 gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa
     2341 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     2401 aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg
     2461 cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag
     2521 gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat ttttctaaat
     2581 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2641 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     2701 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     2761 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     2821 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     2881 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     2941 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3001 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3061 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3121 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     3181 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     3241 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     3301 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     3361 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     3421 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     3481 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     3541 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     3601 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     3661 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3721 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3781 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     3841 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3901 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3961 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4021 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4081 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4141 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4201 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4261 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4321 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     4381 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     4441 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4501 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     4561 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     4621 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     4681 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     4741 cagatcaatt cgcgcgcgaa ggcgaagcgg catgcattta cgttgacacc atcgaatggt
     4801 gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga
     4861 atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg
     4921 tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag
     4981 cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac
     5041 agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg
     5101 tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag
     5161 aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca
     5221 gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct
     5281 gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta
     5341 ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc
     5401 agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg
     5461 gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact
     5521 ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca
     5581 ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt
     5641 ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct
     5701 catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca
     5761 gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc
     5821 ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc
     5881 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     5941 agtgagcgca acgcaattaa tgtaagttag cgcgaattga tctggtttga cagcttatca
     6001 tcgactgcac ggtgcaccaa tgcttctggc gtcaggcagc catcggaagc tgtggtatgg
     6061 ctgtgcaggt cgtaa
//
