LOCUS       sequence-215801-        8000 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 8000)
  TITLE     Salmon Lab plasmids
REFERENCE   2  (bases 1 to 8000)
  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..8000
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             385..642
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    689..814
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1306..1423
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    1783..2016
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2201..2245
                     /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    2522..2542
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    3236..3260
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      3262..3271
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3268..3978
                     /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(3413..3431)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3681..3700
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(3980..4004)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    4022..4610
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4075..4095)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4493..4504)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(4753..4786)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             4806..4986
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5024..5043)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5089..5144
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5143..5162)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5503..5519)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5503..5519)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5516..5538)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5527..5543
                     /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(5551..5557,5558..5575,5576..5581))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5596..5617
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5676..5871
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5722..5857
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5877..6011
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5914..5933)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5968..5987
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6079..6096)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6250..6838)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6330..6349)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7009..7800,7801..7869))
                     /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     7632..7651
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7870..7974)
                     /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 tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggaggcg tggcctgggc gggactgggg agtggcgagc
      421 cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc
      481 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
      541 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
      601 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
      661 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
      721 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
      781 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
      841 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
      901 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
      961 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
     1021 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
     1081 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaaaaa
     1141 agcacagcaa gcagcaggat cttcagacct ggaaattccc tacaatcccc aaagtcaagg
     1201 agtagtagaa tctatgaata aagaattaaa gaaaattata ggacaggtaa gagatcaggc
     1261 tgaacatctt aagacagcag tacaaatggc agtattcatc cacaatttta aaagaaaagg
     1321 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     1381 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     1441 cagcagaaat ccactttgga aaggaccagc aaagctcctc tggaaaggtg aaggggcagt
     1501 agtaatacaa gataatagtg acataaaagt agtgccaaga agaaaagcaa agatcattag
     1561 ggattatgga aaacagatgg caggtgatga ttgtgtggca agtagacagg atgaggatta
     1621 gaacatggaa aagtttagta aaacaccata aggaggagat atgagggaca attggagaag
     1681 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1741 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1801 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1861 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1921 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1981 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     2041 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     2101 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     2161 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     2221 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     2281 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     2341 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     2401 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     2461 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgctagt gattatcgga
     2521 tcaactttgt atagaaaagt tgaccactcc agtggaggct gagaaaggtt ttgtagctgg
     2581 gtagagtatg tactaagaga tggagacggc tggctctgag ctctgaagca agcacctctt
     2641 atggagagtt gctgaccttc aggtgcaaat ctaagatact acaggagaat acaccatggg
     2701 gcttcagccc agttgactcc cgagtgggct atgggtttgt ggaaggagag atagaagaga
     2761 agggaccttt cttcttgaat tctgctttcc ttctacctct gagggtgagc tggggtctca
     2821 gctgaggtga ggacacagct atcagtggga actgtgaaac aacagttcaa gggacaaagt
     2881 tactaggtcc cccaacaact gcagcctcct ggggaatgat gtggaaaaat gctcagccaa
     2941 ggacaaagaa ggcctcaccc tctctgagac aatgtcccct gctgtgaact ggttcatcag
     3001 gccacccagg agcccctatt aagactctaa ttaccctaag gctaagtaga ggtgttgttg
     3061 tccaatgagc actttctgca gacctagcac caggcaagtg tttggaaact gcagcttcag
     3121 cccctctggc catctgctga tccaccctta atgggacaaa cagcaaagtc caggggtcag
     3181 ggggggggtg ctttggacta taaagctagt ggggattcag taacccccag ccctaacaac
     3241 tttgtacaaa aaagcaggct cgccaccatg gtgagcaagg gcgaggagga taacatggcc
     3301 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
     3361 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     3421 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     3481 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     3541 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     3601 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     3661 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     3721 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     3781 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     3841 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
     3901 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     3961 gacgagctgt acaagtaaga cccagctttc ttgtacaaag tggtgataat cgaattccga
     4021 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     4081 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     4141 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     4201 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     4261 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     4321 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     4381 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     4441 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     4501 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     4561 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgggaatt
     4621 cccgcggttc gctttaagac caatgactta caaggcagct gtagatctta gccacttttt
     4681 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atgggatcaa
     4741 ttcaccatgg gaataacttc gtatagcata cattatacga agttatgctg ctttttgctt
     4801 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     4861 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     4921 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     4981 ctagcagcat ctagttgaat cgccagtgtg tcgagctagc agatcttttt ccctctgcca
     5041 aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat
     5101 ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg
     5161 ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg
     5221 ccatatgctg gctgccatga acaaaggtgg ctataaagag gtcatcagta tatgaaacag
     5281 ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta gatttttttt
     5341 atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta
     5401 ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta
     5461 tgaagatccc tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt
     5521 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5581 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5641 tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc agcaaccata
     5701 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     5761 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     5821 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact
     5881 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5941 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     6001 atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6061 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6121 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6181 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6241 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6301 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6361 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6421 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     6481 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6541 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6601 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6661 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     6721 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6781 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6841 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6901 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6961 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7021 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7081 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7141 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7201 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7261 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7321 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7381 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7441 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7501 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7561 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7621 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7681 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7741 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7801 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7861 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7921 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7981 tccccgaaaa gtgccacctg
//
