LOCUS       sequence-218845-        8001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tissue-specific expression of the mCherry living color under the 
            control of the Rat Insulin 2 promoter (RIP).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8001)
  TITLE     Salmon Lab plasmids
REFERENCE   2  (bases 1 to 8001)
  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"
COMMENT     Sequence Label: pCLX-RIP-mCherry
FEATURES             Location/Qualifiers
     source          1..8001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    694..718
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      720..729
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             726..1436
                     /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(871..889)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1139..1158
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(1438..1462)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1480..2068
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1533..1553)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1951..1962)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(2211..2244)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             2264..2444
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(2482..2501)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2547..2602
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2601..2620)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2963..2979)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2963..2979)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2976..2998)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2987..3003
                     /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(3011..3017,3018..3035,3036..3041))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3056..3077
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3136..3331
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3182..3317
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3337..3471
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3374..3393)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3428..3447
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3539..3556)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3710..4298)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3790..3809)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4469..5260,5261..5329))
                     /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     5092..5111
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5330..5434)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5465..5844
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             5846..6103
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6150..6275
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6767..6884
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    7244..7477
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7662..7706
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     protein_bind    7981..8001
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
ORIGIN
        1 accactccag tggaggctga gaaaggtttt gtagctgggt agagtatgta ctaagagatg
       61 gagacagctg gctctgagct ctgaagcaag cacctcttat ggagagttgc tgaccttcag
      121 gtgcaaatct aagatactac aggagaatac accatggggc ttcagcccag ttgactcccg
      181 agtgggctat gggtttgtgg aaggagagat agaagagaag ggacctttct tcttgaattc
      241 tgctttcctt ctacctctga gggtgagctg gggtctcagc tgaggtgagg acacagctat
      301 cagtgggaac tgtgaaacaa cagttcaagg gacaaagtta ctaggtcccc caacaactgc
      361 agcctcctgg ggaatgatgt ggaaaaatgc tcagccaagg acaaagaagg cctcaccctc
      421 tctgagacaa tgtcccctgc tgtgaactgg ttcatcaggc cacccaggag cccctattaa
      481 gactctaatt accctaaggc taagtagagg tgttgttgtc caatgagcac tttctgcaga
      541 cctagcacca ggcaagtgtt tggaaactgc agcttcagcc cctctggcca tctgctgatc
      601 cacccttaat gggacaaaca gcaaagtcca ggggtcaggg ggggggtgct ttggactata
      661 aaactagtgg ggattcagta acccccagcc ctaacaactt tgtacaaaaa agcaggctcg
      721 ccaccatggt gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct
      781 tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
      841 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc
      901 tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga
      961 agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg
     1021 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
     1081 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc
     1141 ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg
     1201 acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg
     1261 acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca
     1321 acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     1381 acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtaagacc
     1441 cagctttctt gtacaaagtg gtgataatcg aattccgata atcaacctct ggattacaaa
     1501 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     1561 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     1621 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     1681 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     1741 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     1801 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     1861 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     1921 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     1981 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     2041 cggatctccc tttgggccgc ctccccgcat cgggaattcc cgcggttcgc tttaagacca
     2101 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
     2161 gggctaattc actcccaacg aagacaagat gggatcaatt caccatggga ataacttcgt
     2221 atagcataca ttatacgaag ttatgctgct ttttgcttgt actgggtctc tctggttaga
     2281 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     2341 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     2401 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagcatct agttgaatcg
     2461 ccagtgtgtc gagctagcag atctttttcc ctctgccaaa aattatgggg acatcatgaa
     2521 gccccttgag catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg
     2581 ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca
     2641 tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa
     2701 caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt
     2761 attccataga aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt
     2821 tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct
     2881 ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc tcgacctgca
     2941 gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3001 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3061 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3121 cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3181 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3241 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3301 aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat
     3361 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3421 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt
     3481 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     3541 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3601 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3661 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3721 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3781 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3841 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3901 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3961 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4021 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4081 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4141 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4201 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4261 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4321 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4381 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4441 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4501 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4561 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4621 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4681 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4741 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     4801 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4861 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     4921 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4981 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5041 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5101 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5161 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5221 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5281 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5341 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5401 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     5461 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
     5521 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     5581 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     5641 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     5701 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     5761 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     5821 gtattagtca tcgctattac catgggaggc gtggcctggg cgggactggg gagtggcgag
     5881 ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac
     5941 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     6001 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     6061 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg
     6121 acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag
     6181 cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg
     6241 gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat
     6301 cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata
     6361 tagtatgggc aagcagggag ctagaacgat tcgcagttaa tactggcctg ttagaaacat
     6421 cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag
     6481 aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga
     6541 taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagaaaa
     6601 aagcacagca agcagcagga tcttcagacc tggaaattcc ctacaatccc caaagtcaag
     6661 gagtagtaga atctatgaat aaagaattaa agaaaattat aggacaggta agagatcagg
     6721 ctgaacatct taagacagca gtacaaatgg cagtattcat ccacaatttt aaaagaaaag
     6781 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     6841 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     6901 acagcagaaa tccactttgg aaaggaccag caaagctcct ctggaaaggt gaaggggcag
     6961 tagtaataca agataatagt gacataaaag tagtgccaag aagaaaagca aagatcatta
     7021 gggattatgg aaaacagatg gcaggtgatg attgtgtggc aagtagacag gatgaggatt
     7081 agaacatgga aaagtttagt aaaacaccat aaggaggaga tatgagggac aattggagaa
     7141 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
     7201 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
     7261 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg
     7321 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
     7381 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
     7441 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
     7501 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
     7561 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
     7621 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
     7681 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
     7741 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
     7801 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
     7861 tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag
     7921 acagagacag atccattcga ttagtgaacg gatctcgacg gtatctagtg attatcggat
     7981 caactttgta tagaaaagtt g
//
