LOCUS       sequence-458395-        6818 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mScarletI-T2A-mTq2 in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6818)
  AUTHORS   Botman D, Tielman A, Goedhart J, Teusink B
  TITLE     An mTurquoise2-based glucose biosensor
  JOURNAL   bioRxiv 2024.11.29.626064
REFERENCE   2  (bases 1 to 6818)
  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: "bioRxiv 
            2024.11.29.626064"
COMMENT     Sequence Label: mScarletI-T2A-mTq2 pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..6818
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(758..776)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1026..1045
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1327..1380
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      1396..1405
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1402..1404,1405..1407,1408..2121)
                     /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(1447..1468)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1708..1727)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2055..2076
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(2137..2155)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2138..2155)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2175..2192)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2181..2405
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2451..2879
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2538..2557)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2748..2769
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2888..2908)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2893..3222
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3073..3208
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3135..3154
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3289..4083
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3343..3362)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3953..3972
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4257..4378
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4294..4313)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4348..4367
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4427..4443)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4427..4443)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4440..4462)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4451..4467
                     /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)."
     promoter        complement(join(4475..4481,4482..4499,4500..4505))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4520..4541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4658..4675)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4829..5417)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4909..4928)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5588..6379,6380..6448))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6211..6230
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6449..6553)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6628..6647)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 atgcggttca aggtgcacat ggagggctcc atgaacggcc acgagttcga gatcgagggc
      721 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
      781 ggccccctgc ccttctcctg ggacatcctg tcccctcagt tcatgtacgg ctccagggcc
      841 ttcatcaagc accccgccga catccccgac tactataagc agtccttccc cgagggcttc
      901 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgccg tgaccgtgac ccaggacacc
      961 tccctggagg acggcaccct gatctacaag gtgaagctcc gcggcaccaa cttccctcct
     1021 gacggccccg taatgcagaa gaagacaatg ggctgggaag cgtccaccga gcggttgtac
     1081 cccgaggacg gcgtgctgaa gggcgacatt aagatggccc tgcgcctgaa ggacggcggc
     1141 cgctacctgg cggacttcaa gaccacctac aaggccaaga agcccgtgca gatgcccggc
     1201 gcctacaacg tcgaccgcaa gttggacatc acctcccaca acgaggacta caccgtggtg
     1261 gaacagtacg aacgctccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     1321 tccggagagg gcagaggaag tcttctaaca tgcggtgacg tggaggagaa tcccggccct
     1381 ggggatccac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     1441 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1501 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1561 ctgcccgtgc cctggcccac cctcgtgacc accctgtcct ggggcgtgca gtgcttcgcc
     1621 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1681 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1741 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1801 gacggcaaca tcctggggca caagctggag tacaactact ttagcgacaa cgtctatatc
     1861 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
     1921 gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1981 gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac
     2041 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     2101 atggacgagc tgtacaagta atctagaggg ccctattcta tagtgtcacc taaatgctag
     2161 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     2221 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     2281 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     2341 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     2401 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     2461 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2521 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2581 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2641 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     2701 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2761 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     2821 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     2881 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     2941 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3001 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3061 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3121 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     3181 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     3241 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     3301 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3361 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3421 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3481 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3541 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3601 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3661 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     3721 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     3781 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     3841 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     3901 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     3961 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4021 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4081 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     4141 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4201 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     4261 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4321 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4381 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     4441 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4501 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4561 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4621 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4681 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4741 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4801 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4861 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4921 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4981 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5041 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5101 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5161 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5221 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5281 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5341 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5401 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5461 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5521 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5581 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5641 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5701 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5761 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5821 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5881 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5941 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6001 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6061 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6121 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6181 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6241 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6301 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6361 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6421 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6481 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6541 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     6601 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     6661 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     6721 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     6781 tgcgctgctt cgcgatgtac gggccagata tacgcgtt
//
