LOCUS       sequence-242466-        5816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cytoplasmic mNeongreen superfolder mTurquoise2 tandem, ~60 % energy 
            transfer. pNM044 - pSAV057-mNG-sfTq2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5816)
  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 5816)
  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: mNG-sfTq2
FEATURES             Location/Qualifiers
     source          1..5816
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1186..1191,1192..1208,1209..1215)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1223..1239
                     /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     1228..1250
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             join(1947..1949,1950..1952,1953..2663)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNVEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1992..2013)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2600..2621
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2684..2713
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2729..2746
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2802..2819)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2972..3058
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3150..3177
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3248..3267)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3249..3267)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3274..3291)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3274..3290)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3283..3305)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        3766..3868
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3869..4528
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3935..3954)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      4589..5134
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     5035..5054
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5251..5268
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5510..5532
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        5610..5687
                     /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(5688..5816,1..954)
                     /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(5707..5726)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg
       61 attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt
      121 aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc
      181 gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc
      241 attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt
      301 ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat
      361 gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg
      421 ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa
      481 tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg
      541 tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg
      601 gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc
      661 gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc
      721 ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc
      781 ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg
      841 gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc
      901 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
      961 cgcaattaat gtaagttagc gcgaattgat ctggtttgac agcttatcat cgactgcacg
     1021 gtgcaccaat gcttctggcg tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc
     1081 gtaaatcact gcataattcg tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt
     1141 gcgccgacat cataacggtt ctggcaaata ttctgaaatg agctgtttac aattaatcat
     1201 ccggctcgta taatgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagacca
     1261 tggcctctct cccagcgaca catgagttac acatctttgg ctccatcaac ggtgtggact
     1321 ttgacatggt gggtcagggc accggcaatc caaatgatgg ttatgaggag ttaaacctga
     1381 agtccaccaa gggtgacctc cagttctccc cctggattct ggtccctcat atcgggtatg
     1441 gcttccatca gtacctgccc taccctgacg ggatgtcgcc tttccaggcc gcgatggtag
     1501 atggctccgg ataccaagtc catcgcacaa tgcagtttga agatggtgcc tcccttactg
     1561 ttaactaccg ctacacctac gagggaagcc acatcaaagg agaggcccag gtgaagggga
     1621 ctggtttccc tgctgacggt cctgtgatga ccaactcgct gaccgctgcg gactggtgca
     1681 ggtcgaagaa gacttacccc aacgacaaaa ccatcatcag tacctttaag tggagttaca
     1741 ccactggaaa tggcaagcgc taccggagca ctgcgcggac cacctacacc tttgccaagc
     1801 caatggcggc taactatctg aagaaccagc cgatgtacgt gttccgtaag acggagctca
     1861 agcactccaa gaccgagctc aacttcaagg agtggcaaaa ggcctttacc gatgtgatgg
     1921 gcatggacga gctgtacaag gaattgatgg tgagcaaggg cgaggagctg ttcaccgggg
     1981 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgcgcg
     2041 gcgagggcga gggcgatgcc accaatggca agctgaccct gaagttcatc tgcaccaccg
     2101 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gtcctggggc gtgcagtgct
     2161 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     2221 gctacgtcca ggagcgcacc atcagcttca aggacgacgg cacatacaag acccgcgccg
     2281 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     2341 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctactttagc gacaacgtct
     2401 atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaacg
     2461 tggaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     2521 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccaagctg agcaaagacc
     2581 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     2641 tcggcatgga cgagctgtac aaggaattct aaagcttggg cccgaacaaa aactcatctc
     2701 agaagaggat ctgaatagcg ccgtcgacca tcatcatcat catcattgag tttaaacggt
     2761 ctccagcttg gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca
     2821 gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca
     2881 cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct
     2941 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     3001 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     3061 gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc
     3121 gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc
     3181 gtttctacaa actcttttgt ttatttttct aaatacattc aaatatgtat ccgctcatgc
     3241 caattcgccc tatagtgagt cgtattacaa ttcactggcc gtcgttttac aacgtcgtga
     3301 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     3361 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     3421 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3481 cagcgtgacc gctacacttg ccagcgctga tgtccggcgg tgcttttgcc gttacgcacc
     3541 accccgtcag tagctgaaca ggagggacag ctgatagaaa cagaagccac tggagcacct
     3601 caaaaacacc atcatacact aaatcagtaa gttggcagca tcacccgacg cactttgcgc
     3661 cgaataaata cctgtgacgg aagatcactt cgcagaataa ataaatcctg gtgtccctgt
     3721 tgataccggg aagccctggg ccaacttttg gcgaaaatga gacgttgatc ggcacgtaag
     3781 aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt ttgagttatc
     3841 gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt
     3901 tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg
     3961 tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa
     4021 taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc
     4081 ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg
     4141 ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga
     4201 cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct
     4261 ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt
     4321 gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt
     4381 cactatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt
     4441 tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta
     4501 ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc agttattggt gcctagaaat
     4561 attttatctg attaataaga tgatcttctt gagatcgttt tggtctgcgc gtaatctctt
     4621 gctctgaaaa cgaaaaaacc gccttgcagg gcggtttttc gaaggttctc tgagctacca
     4681 actctttgaa ccgaggtaac tggcttggag gagcgcagtc accaaaactt gtcctttcag
     4741 tttagcctta accggcgcat gacttcaaga ctaactcctc taaatcaatt accagtggct
     4801 gctgccagtg gtgcttttgc atgtctttcc gggttggact caagacgata gttaccggat
     4861 aaggcgcagc ggtcggactg aacggggggt tcgtgcatac agtccagctt ggagcgaact
     4921 gcctacccgg aactgagtgt caggcgtgga atgagacaaa cgcggccata acagcggaat
     4981 gacaccggta aaccgaaagg caggaacagg agagcgcacg agggagccgc cagggggaaa
     5041 cgcctggtat ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc gtcagatttc
     5101 gtgatgcttg tcaggggggc ggagcctatg gaaaaacggc tttgccgcgg ccctctcact
     5161 tccctgttaa gtatcttcct ggcatcttcc aggaaatctc cgccccgttc gtaagccatt
     5221 tccgctcgcc gcagtcgaac gaccgagcgt agcgagtcag tgagcgagga agcggaatat
     5281 atcctgtatc acatattctg ctgacgcacc ggtgcagcct tttttctcct gccacatgaa
     5341 gcacttcact gacaccctca tcagtgccaa catagtaagc cagtatacac tccgctatcg
     5401 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     5461 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     5521 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagcagat caattcgcgc
     5581 gcgaaggcga agcggcatgc atttacgttg acaccatcga atggtgcaaa acctttcgcg
     5641 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa
     5701 cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga
     5761 accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcg
//
