LOCUS       sequence-375299-        4710 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C1 Cloning vector for mammalian cells incorporating mScarlet-I3 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: pmScarlet-I3_C1
FEATURES             Location/Qualifiers
     source          1..4710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(188..207)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    232..279
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      608..1196
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1097..1116
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1371..1674
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1675..1878
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1829..1849
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1917..1926
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1923..2609
                     /codon_start=1
                     /product="bright cysteine-free monomeric red fluorescent 
                     protein with fast and complete maturation (Gadella et al., 
                     2023)"
                     /label=mScarlet3
                     /translation="MDSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYWKQSFPEGFKWERVMIFEDGGT
                     VSVTQDTSLEDGTLIYKVKLRGGNFPPDGPVMQKRTMGWEASTERLYPEDVVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHS
                     TGGSGGS"
     primer_bind     complement(2053..2071)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_feature    2619..2684
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2808..2929
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2845..2864)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2899..2918
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2936..3391)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3073..3094)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3285..3304
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3418..3522
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3524..3881
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3524..3715
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3547..3567)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3732..3867
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3916..4710
                     /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             3916..4710
                     /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(3970..3989)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4580..4599
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
       61 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
      121 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
      181 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
      241 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
      301 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
      361 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
      421 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
      481 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
      541 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
      601 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
      661 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
      721 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
      781 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
      841 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
      901 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
      961 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     1021 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     1081 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     1141 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     1201 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     1261 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa
     1321 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     1381 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     1441 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     1501 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     1561 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     1621 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     1681 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     1741 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     1801 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     1861 gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggtcgcca
     1921 ccatggatag caccgaggca gtgatcaagg agttcatgcg gttcaaggtg cacatggagg
     1981 gctccatgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     2041 gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc tcctgggaca
     2101 tcctgtcccc tcagttcatg tacggctcca gggccttcat caagcacccc gccgacatcc
     2161 ccgactactg gaagcagtcc ttccccgagg gcttcaagtg ggagcgcgtg atgatcttcg
     2221 aggacggcgg caccgtgtcc gtgacccagg acacctccct ggaggacggc accctgatct
     2281 acaaggtgaa gctccgcggc ggcaacttcc ctcctgacgg ccccgtaatg cagaagagga
     2341 caatgggctg ggaagcatcc accgagcggt tgtaccccga ggacgtcgtg ctgaagggcg
     2401 acattaagat ggccctgcgc ctgaaggacg gcggccgcta cctggcggac ttcaagacca
     2461 cctacaaggc caagaagccc gtgcagatgc ccggcgcctt caacatcgac cgcaagttgg
     2521 acatcacctc ccacaacgag gactacaccg tggtggaaca gtacgaacgc tccgtggccc
     2581 gccactccac cggcggctcc ggtggctcct tgtacaagtc cggactcaga tctcgagctc
     2641 aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg atccaccgga tctagataac
     2701 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     2761 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     2821 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2881 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     2941 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     3001 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     3061 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     3121 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     3181 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     3241 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     3301 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     3361 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     3421 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3481 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     3541 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3601 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3661 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3721 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3781 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3841 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     3901 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3961 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4021 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4081 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4141 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4201 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4261 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4321 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4381 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4441 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4501 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4561 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4621 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4681 ttctatcgcc ttcttgacga gttcttctga
//
