LOCUS       sequence-223034-        5020 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Strong CAG promoter with mCherry expression for transfection 
            efficacy screening.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5020)
  AUTHORS   Mishra B, Wilson DR, Sripathi SR, Suprenant MP, Rui Y, Wahlin KJ, 
            Berlinicke CA, Green JJ, Zack DJ
  TITLE     A combinatorial library of biodegradable polyesters enables 
            non-viral gene delivery to post-mitotic human stem cell-derived 
            polarized RPE monolayers.
  JOURNAL   Regen Eng Transl Med. 2019 Sep;6(3):273-285. doi: 
            10.1007/s40883-019-00118-1. Epub 2019 Jul 24.
  PUBMED    33732871
REFERENCE   2  (bases 1 to 5020)
  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: 
            "10.1007/s40883-019-00118-1"; journalName: "Regen Eng Transl Med"; 
            date: "2019-09"; volume: "6"; issue: "3"; pages: "273-285"
COMMENT     Sequence Label: CAG-mCherry
FEATURES             Location/Qualifiers
     source          1..5020
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     562..584
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     646..665
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    701..725
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             746..1456
                     /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(891..909)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1159..1178
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(1473..1497)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1589..1608)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1654..1709
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1708..1727)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2070..2086)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2070..2086)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2083..2105)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2094..2110
                     /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(2118..2124,2125..2142,2143..2148))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2163..2184
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2243..2438
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2289..2424
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2444..2578
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2481..2500)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2535..2554
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2646..2663)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2817..3405)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2897..2916)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3576..4367,4368..4436))
                     /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     4199..4218
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4437..4541)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4572..4951
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg gggggngcgc
       61 gccggggggg gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg
      121 ggngsggngg gggggggggg ggggggggng gggggggggg gggggggggg gggggggggg
      181 gggggggggg gggggggggg gggggggggs gggggggggg ggggccgcct cgggccgggg
      241 agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga ggcgcggcga
      301 gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc
      361 aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc gggcgcgggg
      421 cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc
      481 gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac ggctgccttc
      541 gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct
      601 ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta
      661 ttgtgctgtc tcatcatttt ggcaaagaat tcctcgaatc acaagtttgt acaaaaaagc
      721 aggctccgcg gccgccccct tcaccatggt gagcaagggc gaggaggata acatggccat
      781 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
      841 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
      901 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
      961 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt
     1021 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
     1081 gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac
     1141 caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc
     1201 cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct
     1261 gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt
     1321 gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga
     1381 ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga
     1441 cgagctgtat aagtaaaagg gtgggcgcgc cgacccagct ttcttgtaca aagtggtgat
     1501 tcgaggaatt cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca
     1561 atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca
     1621 tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa
     1681 tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt
     1741 taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg
     1801 ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc
     1861 attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg
     1921 tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt
     1981 tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg
     2041 acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     2101 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     2161 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     2221 aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc ccgcccctaa
     2281 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2341 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     2401 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc
     2461 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2521 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg
     2581 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2641 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2701 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2761 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2821 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2881 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2941 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3001 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3061 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3121 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3181 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3241 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3301 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3361 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3421 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3481 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3541 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3601 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3661 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3721 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3781 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3841 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3901 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3961 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4021 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4081 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4141 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4201 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4261 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4321 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4381 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4441 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4501 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4561 ccacctgggt cgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     4621 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     4681 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     4741 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     4801 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     4861 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     4921 catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac
     4981 tctccccatc tcccccccct ccccaccccc aattttgtat
//
