LOCUS       sequence-206711-        3633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression vector for mCarmine, a far red emitting 
            fluorescent protein (excitation: 603; emission: 675).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3633)
  AUTHORS   Fabritius A, Ng D, Kist AM, Erdogan M, Portugues R, Griesbeck O
  TITLE     Imaging-Based Screening Platform Assists Protein Engineering.
  JOURNAL   Cell Chem Biol. 2018 Dec 20;25(12):1554-1561.e8. doi: 
            10.1016/j.chembiol.2018.08.008. Epub 2018 Sep 13.
  PUBMED    30220597
REFERENCE   2  (bases 1 to 3633)
  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: 
            "10.1016/j.chembiol.2018.08.008"; journalName: "Cell Chem Biol"; 
            date: "2018-12-20- 20"; volume: "25"; issue: "12"; pages: 
            "1554-1561.e8"
COMMENT     Sequence Label: mCarmine in pRSETB
FEATURES             Location/Qualifiers
     source          1..3633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             112..141
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             148..180
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(184..192,193..207)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      214..223
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             220..954
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein with 
                     favorable expression and pKa properties and with peak 
                     emission at 675 nm (Fabritius et al., 2018)"
                     /label=mCarmine
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTQ
                     RIKVVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLKVKQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTETLYPADGGLRGYNT
                     MALKLVGGGHLQTFLKTTYRSKKPAKNLKMPGVYFVDRRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     primer_bind     complement(967..984)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1034..1052)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1048..1095
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1192..1647
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1279..1298)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1489..1510
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1673..1777
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1778..1846,1847..2638)
                     /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     complement(1996..2015)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2809..3397
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3298..3317
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3551..3568
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat
      121 catcatcacc atcaccatca ccgcggtatg gctagcatga ctggtggaca gcaaatgggt
      181 cgggatctgt acgacgatga cgataaggat ccggccacca tggtgtctaa gggcgaagag
      241 ctgattaagg agaacatgca catgaagctg tacatggagg gcaccgtgaa caaccaccac
      301 ttcaagtgca catccgaggg cgaaggcaag ccctacgagg gcacccagac ccagagaatc
      361 aaggtggtcg agggcggccc tctccccttc gccttcgaca tcctggctac cagcttcatg
      421 tacggcagca agaccttcat caaccacacc cagggcatcc ccgatttctt taagcagtcc
      481 ttccctgagg gcttcacatg ggagagagtc accacatacg aagacggggg cgtgctgaag
      541 gtgaaacagg acaccagcct ccaggacggc tgcctcatct acaacgtcaa gatcagaggg
      601 gtgaacttcc catccaacgg ccctgtgatg cagaagaaaa cactcggctg ggaggccaac
      661 accgagaccc tgtaccccgc tgacggcggc ctgcgcggct ataacacgat ggccctgaag
      721 ctcgtgggcg ggggccacct gcagacgttc ctcaagacca catacagatc caagaaaccc
      781 gctaagaacc tcaagatgcc cggcgtctac tttgtggacc gcagactgga aagaatcaag
      841 gaggccgaca aagagaccta cgtcgagcag cacgaggtgg ctgtggccag atactgcgac
      901 ctccctagca aactggggca caaacttaat ggcatggacg agctgtacaa gtgataatag
      961 gaattcctat agtgtcacct aaatgatccg gctgctaaca aagcccgaaa ggaagctgag
     1021 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     1081 ttgaggggtt ttttgctgaa aggaggaact atatccggat ctggcgtaat agcgaagagg
     1141 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct
     1201 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     1261 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     1321 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     1381 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     1441 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     1501 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     1561 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     1621 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
     1681 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     1741 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     1801 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     1861 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     1921 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     1981 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     2041 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     2101 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     2161 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     2221 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     2281 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     2341 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     2401 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     2461 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     2521 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     2581 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     2641 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     2701 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     2761 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2821 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2881 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2941 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3001 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3061 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3121 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3181 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3241 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3301 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3361 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3421 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     3481 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     3541 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     3601 tctccccgcg cgttggccga ttcattaatg cag
//
