LOCUS       sequence-242470-        6226 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Periplasmic OM bound mNeongreen superfolder mTurquoise2 ox tandem, 
            ~40 % energy transfer. pNM092 - 
            pTHV037-Ompa-SA1-177-(SA-1)-LEDPPAEL-mNG-sfTq2ox, .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6226)
  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 6226)
  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: OmpA-mNG-sfTq2ox
FEATURES             Location/Qualifiers
     source          1..6226
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(53..58,59..75,76..82)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    90..106
                     /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     95..117
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     sig_peptide     133..192
                     /codon_start=1
                     /product="signal peptide from the E. coli outer membrane 
                     protein OmpA"
                     /label=OmpA signal peptide
                     /translation="KKTAIAIAVALAGFATVAQA"
     CDS             793..1500
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     CDS             join(1507..1509,1510..1512,1513..2223)
                     /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(1552..1573)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2160..2181
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2270..2287)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2440..2526
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2618..2645
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2664..2755
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2756..2824,2825..3616)
                     /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(2974..2993)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3787..4375
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4276..4295
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4529..4546
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4561..4701
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4627..4646)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4787..4809
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4887..4964
                     /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             4965..6047
                     /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(4984..5003)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc tgtttacaat
       61 taatcatccg gctcgtataa tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
      121 cagaccatgg caaaaaagac agctatcgcg attgcagtgg cactggctgg tttcgctacc
      181 gtagcgcagg ccgctccgaa agataacacc tggtacactg gtgctaaact gggctggtcc
      241 cagtaccatg acactggcct catcggccaa cgcttggaga tctgccagaa cgtttgctac
      301 tacctcggga cgctcaatgg tccgacccat gaaaaccaac tgggcgctgg tgcttttggt
      361 ggttaccagg ttaacccgta tgttggcttt gaaatgggtt acgactggtt aggtcgtatg
      421 ccgtacaaag gcagcgttga aaacggtgca tacaaagctc agggcgttca actgaccgct
      481 aaactgggtt acccaatcac tgacgacctg gacatctaca ctcgtctggg tggcatggta
      541 tggcgtgcag acactaaatc caacgtttat ggtaaaaacc acgacaccgg cgtttctccg
      601 gtcttcgctg gcggtgttga gtacgcgatc actcctgaaa tcgctacccg tctggaatac
      661 cagtggacca acaacatcgg tgacgcaggc cagatcggcc agcgtccgga caacggcatg
      721 ctgagcctgg gtgtttccta ccgtttcggt cagggcgaag cagctccact tgaggatccc
      781 cccgctgaat tgatggtgag caagggcgag gaggataaca tggcctctct cccagcgaca
      841 catgagttac acatctttgg ctccatcaac ggtgtggact ttgacatggt gggtcagggc
      901 accggcaatc caaatgatgg ttatgaggag ttaaacctga agtccaccaa gggtgacctc
      961 cagttctccc cctggattct ggtccctcat atcgggtatg gcttccatca gtacctgccc
     1021 taccctgacg ggatgtcgcc tttccaggcc gcgatggtag atggctccgg ataccaagtc
     1081 catcgcacaa tgcagtttga agatggtgcc tcccttactg ttaactaccg ctacacctac
     1141 gagggaagcc acatcaaagg agaggcccag gtgaagggga ctggtttccc tgctgacggt
     1201 cctgtgatga ccaactcgct gaccgctgcg gactggtgca ggtcgaagaa gacttacccc
     1261 aacgacaaaa ccatcatcag tacctttaag tggagttaca ccactggaaa tggcaagcgc
     1321 taccggagca ctgcgcggac cacctacacc tttgccaagc caatggcggc taactatctg
     1381 aagaaccagc cgatgtacgt gttccgtaag acggagctca agcactccaa gaccgagctc
     1441 aacttcaagg agtggcaaaa ggcctttacc gatgtgatgg gcatggacga gctgtacaag
     1501 gaattgatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     1561 ctggacggcg acgtaaacgg ccacaagttc agcgtgcgcg gcgagggcga gggcgatgcc
     1621 accaatggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     1681 cccaccctcg tgaccaccct gtcctggggt gtacaggtct tcgcccgcta ccccgaccac
     1741 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     1801 atcagcttca aggacgacgg cacatacaag acccgcgccg aggtgaagtt cgagggcgac
     1861 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     1921 gggcacaagc tggagtacaa ctactttagc gacaacgtct atatcaccgc cgacaagcag
     1981 aagaacggca tcaaggccaa cttcaagatc cgccacaacg tggaggacgg cggcgtgcag
     2041 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     2101 aaccactacc tgagcaccca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac
     2161 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     2221 aaggaattct aaagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga
     2281 ttaaatcaga acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg
     2341 tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg
     2401 tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag
     2461 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     2521 acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca
     2581 ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc
     2641 ctttttgcgt ttctacaaac tcttttgttt atttttctaa atacattcaa atatgtatcc
     2701 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     2761 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2821 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2881 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2941 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     3001 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3061 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3121 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     3181 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3241 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     3301 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3361 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3421 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3481 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3541 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3601 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3661 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3721 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3781 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3841 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3901 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     3961 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4021 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4081 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4141 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4201 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4261 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4321 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4381 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4441 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4501 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     4561 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac
     4621 tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta
     4681 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4741 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4801 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agcagatcaa ttcgcgcgcg
     4861 aaggcgaagc ggcatgcatt tacgttgaca ccatcgaatg gtgcaaaacc tttcgcggta
     4921 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt
     4981 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
     5041 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
     5101 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
     5161 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
     5221 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
     5281 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
     5341 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
     5401 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg
     5461 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
     5521 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
     5581 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
     5641 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
     5701 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
     5761 cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt
     5821 taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
     5881 aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa
     5941 gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     6001 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     6061 aatgtaagtt agcgcgaatt gatctggttt gacagcttat catcgactgc acggtgcacc
     6121 aatgcttctg gcgtcaggca gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc
     6181 actgcataat tcgtgtcgct caaggcgcac tcccgttctg gataat
//
