LOCUS       sequence-227545-        6075 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 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: sfTq2ox-PBP5
FEATURES             Location/Qualifiers
     source          1..6075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(174..179,180..196,197..203)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    211..227
                     /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     216..238
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             join(344..346,347..349,350..1060)
                     /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(389..410)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     997..1018
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2240..2257)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2410..2496
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2588..2615
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2634..2725
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2726..2794,2795..3586)
                     /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(2944..2963)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3757..4345
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4246..4265
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4499..4516
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4531..4671
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4597..4616)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4757..4779
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4857..4934
                     /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             4935..6017
                     /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(4954..4973)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 actgcacggt gcaccaatgc ttctggcgtc aggcagccat cggaagctgt ggtatggctg
       61 tgcaggtcgt aaatcactgc ataattcgtg tcgctcaagg cgcactcccg ttctggataa
      121 tgttttttgc gccgacatca taacggttct ggcaaatatt ctgaaatgag ctgtttacaa
      181 ttaatcatcc ggctcgtata atgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
      241 acagaccatg ggcaaaaaga tttggctggc gctggctggt ttagttttag cgtttagcgc
      301 atcggcggcg cagtatgaag atctcgaggg tccggctggt ctgatggtga gcaagggcga
      361 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
      421 caagttcagc gtgcgcggcg agggcgaggg cgatgccacc aatggcaagc tgaccctgaa
      481 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgtc
      541 ctggggtgta caggtcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
      601 gtccgccatg cccgaaggct acgtccagga gcgcaccatc agcttcaagg acgacggcac
      661 atacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
      721 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
      781 ctttagcgac aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt
      841 caagatccgc cacaacgtgg aggacggcgg cgtgcagctc gccgaccact accagcagaa
      901 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
      961 caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1021 cgccgccggg atcactctcg gcatggacga gctgtacaag gaattcggat ccgatgacct
     1081 gaatatcaaa actatgatcc cgggtgtacc gcagatcgat gcggagtcct acatcctgat
     1141 tgactataac tccggcaaag tgctcgccga acagaacgca gatgtccgcc gcgatcctgc
     1201 cagcctgacc aaaatgatga ccagttacgt tatcggccag gcaatgaaag ccggtaaatt
     1261 taaagaaact gatttagtca ctatcggcaa cgacgcatgg gccaccggta acccggtgtt
     1321 taaaggttct tcgctgatgt tcctcaaacc gggcatgcag gttccggttt ctcagctgat
     1381 ccgcggtatt aacctgcaat cgggtaacga tgcttgtgtc gccatggccg attttgccgc
     1441 tggtagccag gacgcttttg ttggcttgat gaacagctac gttaacgcac tgggcctgaa
     1501 aaatacccac ttccagacgg tacatggtct ggatgctgat ggtcagtaca gctccgcgcg
     1561 agatatggcg ctgatcggcc aggcattgat ccgtgacgta ccgaatgaat actcgatcta
     1621 taaagaaaaa gaatttacgt ttaacggtat tcgccagctg aaccgtaacg gcctgttatg
     1681 ggataacagc ctgaatgtcg acggcatcaa aaccggacac actgacaaag caggttacaa
     1741 ccttgttgct tctgcgactg aaggccagat gcgcttgatt tctgcggtaa tgggcggacg
     1801 tacttttaaa ggccgtgaag ccgaaagtaa aaaactgcta acctggggct tccgtttctt
     1861 tgaaaccgtt aacccactga aagtaggtaa agagttcgcc tctgaaccgg tttggtttgg
     1921 tgattctgat cgcgcttcgt taggggttga taaagacgtg tacctgacca ttccgcgtgg
     1981 tcgcatgaaa gatctgaaag ccagctatgt gctgaacagc agtgaattgc atgcgccgct
     2041 gcaaaagaat caggtcgtcg gaactatcaa cttccagctt gatggcaaaa cgatcgagca
     2101 acgcccgctg gttgtgttgc aagaaatccc ggaaggtaac ttcttcggca aaatcattga
     2161 ttacattaaa ttaatgttcc atcactggtt tggttaaaaa gaagcttggc tgttttggcg
     2221 gatgagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa
     2281 aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag
     2341 aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact
     2401 gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     2461 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
     2521 gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa
     2581 attaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac tcttttgttt
     2641 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     2701 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     2761 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     2821 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     2881 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     2941 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     3001 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     3061 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     3121 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     3181 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     3241 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     3301 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     3361 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     3421 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     3481 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     3541 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     3601 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     3661 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     3721 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3781 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3841 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3901 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3961 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4021 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4081 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4141 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4201 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4261 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4321 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4381 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     4441 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     4501 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     4561 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     4621 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     4681 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4741 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4801 cgcgcgaggc agcagatcaa ttcgcgcgcg aaggcgaagc ggcatgcatt tacgttgaca
     4861 ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat
     4921 tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct
     4981 cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg
     5041 aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac
     5101 tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc
     5161 cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg
     5221 tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg
     5281 cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg
     5341 tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca
     5401 tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg
     5461 cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc
     5521 tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac
     5581 gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg
     5641 gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg
     5701 ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata
     5761 ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc
     5821 tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca
     5881 atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa
     5941 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     6001 tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agcgcgaatt gatctggttt
     6061 gacagcttat catcg
//
