LOCUS       sequence-435943-        4784 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express a protein of interest fused to the C-terminus of 
            mScarlet3 (unpublished).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4784)
  TITLE     Mammalian expression empty backbones
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4784)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pCS2+mScarlet3-C Cloning Vector
FEATURES             Location/Qualifiers
     source          1..4784
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             78..764
                     /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     476..495
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    836..970
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(860..879)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     914..933
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1083..1103)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1083..1101)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1122..1138)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1122..1138)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1135..1157)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1146..1162
                     /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(1170..1176,1177..1194,1195..1200))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1215..1236
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1353..1370)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1524..2112)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1604..1623)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2283..3074,3075..3143))
                     /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     2906..2925
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3144..3248)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3274..3729)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3411..3432)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3623..3642
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3800..4784
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agacttgttc
       61 tttttgcagg atctaccatg gatagcaccg aggcagtgat caaggagttc atgcggttca
      121 aggtgcacat ggagggctcc atgaacggcc acgagttcga gatcgagggc gagggcgagg
      181 gccgccccta cgagggcacc cagaccgcca agctgagggt gaccaagggt ggccccctgc
      241 ccttctcctg ggacatcctg tcccctcagt tcatgtacgg ctccagggcc ttcacgaagc
      301 accccgccga catccccgac tactggaagc agtccttccc cgagggcttc aagtgggagc
      361 gcgtgatgaa cttcgaggac ggcggcgccg tgtccgtggc ccaggacacc tccctggagg
      421 acggcaccct gatctacaag gtgaagctcc gcggcaccaa cttccctcct gacggccccg
      481 taatgcagaa gaagacaatg ggctgggaag catccaccga gcggttgtac cccgaggacg
      541 tcgtgctgaa gggcgacatt aagatggccc tgcgcctgaa ggacggcggc cgctacctgg
      601 cggacttcaa gaccacctac agggccaaga agcccgtgca gatgcccggc gccttcaaca
      661 tcgaccgcaa gttggacatc acatcccaca acgaggacta caccgtggtg gaacagtacg
      721 aacgctccgt ggcccgccac tccaccggcg gctccggtgg ctccggatct ggatcccatc
      781 gattcgaatt caaggcctct cgagcctcta gaactatagt gagtcgtatt acgtagatcc
      841 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
      901 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
      961 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     1021 ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg gcccggtacc cagcttttgt
     1081 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     1141 tgaaattgtt atccgctcac aattccacac aacatacgag ccgggagcat aaagtgtaaa
     1201 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     1261 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     1321 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     1381 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     1441 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     1501 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1561 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1621 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1681 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1741 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     1801 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1861 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1921 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     1981 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2041 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2101 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     2161 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     2221 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     2281 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     2341 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     2401 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     2461 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     2521 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     2581 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     2641 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     2701 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     2761 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     2821 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     2881 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     2941 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     3001 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     3061 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     3121 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     3181 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     3241 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
     3301 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     3361 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     3421 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     3481 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     3541 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     3601 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     3661 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     3721 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     3781 cctcttcgct attacgccag tcgaccgcca attcaatatg gcgtatatgg actcatgcca
     3841 attcaatatg gtggatctgg acctgtgcca attcaatatg gcgtatatgg actcgtgcca
     3901 attcaatatg gtggatctgg accccagcca attcaatatg gcggacttgg caccatgcca
     3961 attcaatatg gcggacctgg cactgtgcca actggggagg ggtctacttg gcacggtgcc
     4021 aagtttgagg aggggtcttg gccctgtgcc aagtccgcca tattgaattg gcatggtgcc
     4081 aataatggcg gccatattgg ctatatgcca ggatcaatat ataggcaata tccaatatgg
     4141 ccctatgcca atatggctat tggccaggtt caatactatg tattggccct atgccatata
     4201 gtattccata tatgggtttt cctattgacg tagatagccc ctcccaatgg gcggtcccat
     4261 ataccatata tggggcttcc taataccgcc catagccact cccccattga cgtcaatggt
     4321 ctctatatat ggtctttcct attgacgtca tatgggcggt cctattgacg tatatggcgc
     4381 ctcccccatt gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc attgacgtca
     4441 ataggaccac ccaccattga cgtcaatggg atggctcatt gcccattcat atccgttctc
     4501 acgcccccta ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat caatatctat
     4561 taatagtaac ttggcaagta cattactatt ggaagtacgc cagggtacat tggcagtact
     4621 cccattgacg tcaatggcgg taaatggccc gcgatggctg ccaagtacat ccccattgac
     4681 gtcaatgggg aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc
     4741 gtgcctaatg ggaggtctat ataagcaatg ctcgtttagg gaac
//
