LOCUS       sequence-375291-        4710 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C1 Cloning vector for mammalian cells incorporating mScarlet3 red 
            fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4710)
  AUTHORS   Gadella TWJ Jr, van Weeren L, Stouthamer J, Hink MA, Wolters AHG, 
            Giepmans BNG, Aumonier S, Dupuy J, Royant A
  TITLE     mScarlet3: a brilliant and fast-maturing red fluorescent protein.
  JOURNAL   Nat Methods. 2023 Apr;20(4):541-545. doi: 
            10.1038/s41592-023-01809-y. Epub 2023 Mar 27.
  PUBMED    36973546
REFERENCE   2  (bases 1 to 4710)
  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: 
            "10.1038/s41592-023-01809-y"; journalName: "Nat Methods"; date: 
            "2023-04"; volume: "20"; issue: "4"; pages: "541-545"
COMMENT     Sequence Label: pmScarlet3_C1
FEATURES             Location/Qualifiers
     source          1..4710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             135..929
                     /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             135..929
                     /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(189..208)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     799..818
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1117..1136)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1161..1208
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1537..2125
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2026..2045
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2300..2603
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2604..2807
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2758..2778
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2846..2855
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2852..3538
                     /codon_start=1
                     /product="bright cysteine-free monomeric red fluorescent 
                     protein with fast and complete maturation (Gadella et al., 
                     2023)"
                     /label=mScarlet3
                     /translation="MDSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LRVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYWKQSFPEGFKWERVMNFEDGGA
                     VSVAQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDVVLKGDIKM
                     ALRLKDGGRYLADFKTTYRAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHS
                     TGGSGGS"
     primer_bind     3250..3269
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3548..3613
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3737..3858
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3774..3793)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3828..3847
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3865..4320)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4002..4023)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4214..4233
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4347..4451
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(4453..4710,1..100)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4453..4644
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4476..4496)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(4661..4710,1..86)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
       61 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
      121 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
      181 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
      241 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
      301 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
      361 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
      421 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
      481 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
      541 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
      601 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
      661 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
      721 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
      781 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
      841 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
      901 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
      961 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     1021 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     1081 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     1141 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     1201 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     1261 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     1321 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     1381 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     1441 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     1501 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     1561 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     1621 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     1681 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     1741 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     1801 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     1861 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     1921 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     1981 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2041 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     2101 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     2161 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     2221 ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata
     2281 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     2341 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     2401 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     2461 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     2521 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     2581 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
     2641 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     2701 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     2761 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
     2821 gtcagatccg ctagcgctac cggtcgccac catggatagc accgaggcag tgatcaagga
     2881 gttcatgcgg ttcaaggtgc acatggaggg ctccatgaac ggccacgagt tcgagatcga
     2941 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga gggtgaccaa
     3001 gggtggcccc ctgcccttct cctgggacat cctgtcccct cagttcatgt acggctccag
     3061 ggccttcacg aagcaccccg ccgacatccc cgactactgg aagcagtcct tccccgaggg
     3121 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gccgtgtccg tggcccagga
     3181 cacctccctg gaggacggca ccctgatcta caaggtgaag ctccgcggca ccaacttccc
     3241 tcctgacggc cccgtaatgc agaagaagac aatgggctgg gaagcatcca ccgagcggtt
     3301 gtaccccgag gacgtcgtgc tgaagggcga cattaagatg gccctgcgcc tgaaggacgg
     3361 cggccgctac ctggcggact tcaagaccac ctacagggcc aagaagcccg tgcagatgcc
     3421 cggcgccttc aacatcgacc gcaagttgga catcacatcc cacaacgagg actacaccgt
     3481 ggtggaacag tacgaacgct ccgtggcccg ccactccacc ggcggctccg gtggctcctt
     3541 gtacaagtcc ggactcagat ctcgagctca agcttcgaat tctgcagtcg acggtaccgc
     3601 gggcccggga tccaccggat ctagataact gatcataatc agccatacca catttgtaga
     3661 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     3721 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3781 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3841 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     3901 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     3961 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     4021 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     4081 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     4141 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     4201 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     4261 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     4321 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4381 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4441 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     4501 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4561 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4621 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4681 cagttccgcc cattctccgc cccatggctg
//
