LOCUS       sequence-477198-        4078 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4078)
  TITLE     CTRL Enables Gene-specific RNA Regulation Using CRISPR-Cas7-11
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4078)
  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            "Unpublished"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: 1162BY-CMV-mCherry-bGHpolyA
FEATURES             Location/Qualifiers
     source          1..4078
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        36..415
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        416..619
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     570..590
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             661..1371
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(806..824)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1074..1093
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1377..1394
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1417..1434)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1423..1647
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1718..1734)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1718..1734)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1731..1753)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1742..1758
                     /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(1766..1772,1773..1790,1791..1796))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1811..1832
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1949..1966)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2120..2708)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2200..2219)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2879..3670,3671..3739))
                     /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     3502..3521
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3740..3844)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3923..3942)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
       61 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      121 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      181 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      241 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      301 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      361 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      421 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      481 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      541 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      601 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagagat ccgcggccgc
      661 atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag
      721 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
      781 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
      841 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac
      901 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc
      961 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
     1021 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
     1081 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc
     1141 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct
     1201 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc
     1261 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
     1321 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta accggtcatc
     1381 atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     1441 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     1501 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     1561 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     1621 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcga
     1681 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     1741 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     1801 aggatgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     1861 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg ggaagaggcg
     1921 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     1981 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2041 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2101 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2161 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2221 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2281 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2341 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2401 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2461 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2521 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2581 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2641 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2701 gatctcaaga agatcctttg atcttttcta cggggtctga cactcagtgg aacgaaaact
     2761 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     2821 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     2881 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     2941 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3001 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3061 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3121 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3181 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3241 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3301 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3361 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3421 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     3481 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     3541 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     3601 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     3661 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     3721 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     3781 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     3841 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat cgatctcccg
     3901 atcccctagg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     3961 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     4021 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgc
//
