LOCUS       sequence-232038-        5529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of HcRed.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5529)
  AUTHORS   Matsuda T, Cepko CL
  TITLE     Electroporation and RNA interference in the rodent retina in vivo 
            and in vitro.
  JOURNAL   Proc Natl Acad Sci U S A. 2004 Jan 6. 101(1):16-22.
  PUBMED    14603031
REFERENCE   2  (bases 1 to 5529)
  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: "Proc Natl 
            Acad Sci U S A. 2004 Jan 6. 101(1):16-22."
COMMENT     Sequence Label: pCAG-HcRed
FEATURES             Location/Qualifiers
     source          1..5529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..660
                     /label=chicken beta-actin promoter
     intron          661..1677
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1601..1623
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1685..1704
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1772..1781
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1778..2464
                     /codon_start=1
                     /product="far-red fluorescent variant of Heteractis crispa 
                     chromoprotein"
                     /label=HcRed1
                     /note="mammalian codon-optimized"
                     /translation="MVSGLLKESMRIKMYMEGTVNGHYFKCEGEGDGNPFAGTQSMRIH
                     VTEGAPLPFAFDILAPCCEYGSRTFVHHTAEIPDFFKQSFPEGFTWERTTTYEDGGILT
                     AHQDTSLEGNCLIYKVKVHGTNFPADGPVMKNKSGGWEPSTEVVYPENGVLCGRNVMAL
                     KVGDRHLICHHYTSYRSKKAVRALTMPGFHFTDIRLQMLRKKKDEYFELYEASVARYSD
                     LPEKAN"
     primer_bind     complement(2551..2570)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2616..2671
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2670..2689)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3032..3048)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3032..3048)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3045..3067)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3056..3072
                     /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(3080..3086,3087..3104,3105..3110))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3125..3146
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3205..3400
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3251..3386
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3406..3540
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3443..3462)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3497..3516
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3608..3625)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3779..4367)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3859..3878)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4538..5329,5330..5398))
                     /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     5161..5180
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5399..5503)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggcgcgcg ccrggggggg gggggggggg sggggggsgg
      541 ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
      601 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
      661 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
      721 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
      781 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
      841 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc
      901 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc
      961 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg
     1021 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     1081 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac
     1141 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt
     1201 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg
     1261 gggccgcctc gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc
     1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
     1381 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
     1441 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
     1501 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
     1561 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
     1621 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
     1681 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt ctgcagtcga
     1741 cggtaccccg cgggcccggg atccaccggt cgccaccatg gtgagcggcc tgctgaagga
     1801 gagtatgcgc atcaagatgt acatggaggg caccgtgaac ggccactact tcaagtgcga
     1861 gggcgagggc gacggcaacc ccttcgccgg cacccagagc atgagaatcc acgtgaccga
     1921 gggcgccccc ctgcccttcg ccttcgacat cctggccccc tgctgcgagt acggcagcag
     1981 gaccttcgtg caccacaccg ccgagatccc cgacttcttc aagcagagct tccccgaggg
     2041 cttcacctgg gagagaacca ccacctacga ggacggcggc atcctgaccg cccaccagga
     2101 caccagcctg gagggcaact gcctgatcta caaggtgaag gtgcacggca ccaacttccc
     2161 cgccgacggc cccgtgatga agaacaagag cggcggctgg gagcccagca ccgaggtggt
     2221 gtaccccgag aacggcgtgc tgtgcggccg gaacgtgatg gccctgaagg tgggcgaccg
     2281 gcacctgatc tgccaccact acaccagcta ccggagcaag aaggccgtgc gcgccctgac
     2341 catgcccggc ttccacttca ccgacatccg gctccagatg ctgcggaaga agaaggacga
     2401 gtacttcgag ctgtacgagg ccagcgtggc ccggtacagc gacctgcccg agaaggccaa
     2461 ctgaagcggc cgcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc
     2521 caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga
     2581 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc
     2641 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     2701 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc
     2761 tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt
     2821 ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt
     2881 tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac tagccagatt
     2941 tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct
     3001 cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     3061 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     3121 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     3181 gaaacctgtc gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct
     3241 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3301 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     3361 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctaactt gtttattgca
     3421 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3481 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     3541 cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     3601 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3661 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3721 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3781 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3841 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3901 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3961 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4021 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4081 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4141 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4201 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4261 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4321 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4381 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4441 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4501 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4561 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4621 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4681 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4741 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4801 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4861 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4921 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4981 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5041 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5101 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5161 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5221 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5281 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5341 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5401 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5461 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5521 tgccacctg
//
