LOCUS       sequence-210811-        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     19..38
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        19..37
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             69..91
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             111..140
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             147..179
                     /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(183..191,192..206)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      213..222
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             219..953
                     /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(966..983)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1033..1051)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1047..1094
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1191..1646
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1278..1297)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1488..1509
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1672..1776
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1777..1845,1846..2637)
                     /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(1995..2014)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2808..3396
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3297..3316
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3550..3567
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta
       61 gaaataattt tgtttaactt taagaaggag atatacatat gcggggttct catcatcatc
      121 atcatcacca tcaccatcac cgcggtatgg ctagcatgac tggtggacag caaatgggtc
      181 gggatctgta cgacgatgac gataaggatc cggccaccat ggtgtctaag ggcgaagagc
      241 tgattaagga gaacatgcac atgaagctgt acatggaggg caccgtgaac aaccaccact
      301 tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc cagagaatca
      361 aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctacc agcttcatgt
      421 acggcagcaa gaccttcatc aaccacaccc agggcatccc cgatttcttt aagcagtcct
      481 tccctgaggg cttcacatgg gagagagtca ccacatacga agacgggggc gtgctgaagg
      541 tgaaacagga caccagcctc caggacggct gcctcatcta caacgtcaag atcagagggg
      601 tgaacttccc atccaacggc cctgtgatgc agaagaaaac actcggctgg gaggccaaca
      661 ccgagaccct gtaccccgct gacggcggcc tgcgcggcta taacacgatg gccctgaagc
      721 tcgtgggcgg gggccacctg cagacgttcc tcaagaccac atacagatcc aagaaacccg
      781 ctaagaacct caagatgccc ggcgtctact ttgtggaccg cagactggaa agaatcaagg
      841 aggccgacaa agagacctac gtcgagcagc acgaggtggc tgtggccaga tactgcgacc
      901 tccctagcaa actggggcac aaacttaatg gcatggacga gctgtacaag tgataatagg
      961 aattcctata gtgtcaccta aatgatccgg ctgctaacaa agcccgaaag gaagctgagt
     1021 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
     1081 tgaggggttt tttgctgaaa ggaggaacta tatccggatc tggcgtaata gcgaagaggc
     1141 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     1201 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     1261 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     1321 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     1381 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     1441 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     1501 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     1561 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     1621 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     1681 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     1741 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     1801 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     1861 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     1921 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     1981 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     2041 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     2101 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     2161 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     2221 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2281 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     2341 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     2401 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     2461 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     2521 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     2581 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     2641 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     2701 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     2761 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     2821 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     2881 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     2941 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3001 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3061 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3121 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3181 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3241 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3301 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3361 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3421 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     3481 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     3541 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     3601 ctccccgcgc gttggccgat tcattaatgc agg
//
