LOCUS       sequence-191170-        5399 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces a fusion between mTurquoise2 and mNeonGreen. It can be used
            as a positive control for FRET.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5399)
  AUTHORS   Mastop M, Bindels DS, Shaner NC, Postma M, Gadella TWJ Jr, Goedhart 
            J
  TITLE     Characterization of a spectrally diverse set of fluorescent proteins
            as FRET acceptors for mTurquoise2.
  JOURNAL   Sci Rep. 2017 Sep 20;7(1):11999. doi: 10.1038/s41598-017-12212-x.
  PUBMED    28931898
REFERENCE   2  (bases 1 to 5399)
  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"; journalName: "Sci Rep."; 
            date: "2017-09-20"; volume: "7(1):11999. doi"; pages: " 
            10.1038/s41598-017-12212-x"
COMMENT     Sequence Label: mNeonGreen-mTurquoise2
FEATURES             Location/Qualifiers
     source          1..5399
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        44..235
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     192..212
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(237..341)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      368..823
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(455..474)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     665..686
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    830..951
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(841..860)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     895..914
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(1074..1787,1788..1790,1791..1793))
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1119..1140)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1468..1487
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1727..1748
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1790..1799
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1830..2537)
                     /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"
     regulatory      2273..2282
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2534..2543
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2570..2773)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2599..2619)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2774..3077
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3252..3840)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3332..3351)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4169..4216
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4241..4260
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4448..5242)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4448..5242)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4559..4578)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5169..5188
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5277..5399,1..235))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5291..5399,1..27)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
       61 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
      121 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
      181 ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc ctcaggactc
      241 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
      301 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
      361 ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
      421 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
      481 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
      541 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
      601 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
      661 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
      721 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
      781 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttacgcctt aagatacatt
      841 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
      901 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
      961 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     1021 taaaacctct acaaatgtgg tatggctgat tatgatctag agtcgcggcc gctttacttg
     1081 tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag caggaccatg
     1141 tgatcgcgct tctcgttggg gtctttgctc agcttggact gggtgctcag gtagtggttg
     1201 tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc
     1261 tgcacgccgc cgtcctcgat gttgtggcgg atcttgaagt tggccttgat gccgttcttc
     1321 tgcttgtcgg cggtgatata gacgttgtcg ctaaagtagt tgtactccag cttgtgcccc
     1381 aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg
     1441 tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg
     1501 gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg
     1561 tggtcggggt agcgggcgaa gcactgcacg ccccaggaca gggtggtcac gagggtgggc
     1621 cagggcacgg gcagcttgcc ggtggtgcag atgaacttca gggtcagctt gccgtaggtg
     1681 gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc
     1741 tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac catggtggcg
     1801 accggtggat ccgaagatct gagtccggac ttgtacagct cgtccatgcc catcacatcg
     1861 gtaaaggcct tttgccactc cttgaagttg agctcggtct tggagtgctt gagctccgtc
     1921 ttacggaaca cgtacatcgg ctggttcttc agatagttag ccgccattgg cttggcaaag
     1981 gtgtaggtgg tccgcgcagt gctccggtag cgcttgccat ttccagtggt gtaactccac
     2041 ttaaaggtac tgatgatggt tttgtcgttg gggtaagtct tcttcgacct gcaccagtcc
     2101 gcagcggtca gcgagttggt catcacagga ccgtcagcag ggaaaccagt ccccttcacc
     2161 tgggcctctc ctttgatgtg gcttccctcg taggtgtagc ggtagttaac agtaagggag
     2221 gcaccatctt caaactgcat tgtgcgatgg acttggtagc cggagccatc taccatggcg
     2281 gcctggaaag gcgacatccc gtcagggtag ggcaggtact gatggaagcc atacccgata
     2341 tgagggacca gaatccaggg ggagaactgg aggtcaccct tggtggactt caggtttaac
     2401 tcctcataac catcatttgg attgccggtg ccctgaccca ccatgtcaaa gtccacaccg
     2461 ttgatggagc caaagatgtg taactcatgt gtcgctggga gagaggccat gttatcctcc
     2521 tcgcccttgc tcaccatggt ggctagcgga tctgacggtt cactaaacca gctctgctta
     2581 tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac
     2641 gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg
     2701 ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc
     2761 aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa
     2821 agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt
     2881 caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc
     2941 gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat
     3001 acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta
     3061 ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg
     3121 attactatta ataactaatg catggcggta atacggttat ccacagaatc aggggataac
     3181 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3241 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3301 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3361 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3421 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3481 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3541 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3601 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3661 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3721 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3781 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3841 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3901 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3961 tgaagtttta aatcaatcta aagtatatat gagtaacctg aggctatggc agggcctgcc
     4021 gccccgacgt tggctgcgag ccctgggcct tcacccgaac ttggggggtg gggtggggaa
     4081 aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg gtggggtatc gacagagtgc
     4141 cagccctggg accgaacccc gcgtttatga acaaacgacc caacaccgtg cgttttattc
     4201 tgtcttttta ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca
     4261 gttagcctcc ccctagggtg ggcgaagaac tccagcatga gatccccgcg ctggaggatc
     4321 atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa ggcggcggtg
     4381 gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc gaaccccaga
     4441 gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag
     4501 cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa
     4561 tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt
     4621 cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat
     4681 gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg
     4741 ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca
     4801 tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg
     4861 gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag
     4921 caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc
     4981 ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg
     5041 atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa
     5101 aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg
     5161 tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt
     5221 gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcgatctttg
     5281 caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag
     5341 gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggc
//
