LOCUS       sequence-384388-        7514 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hipp11 targeting vector, expresses mScarlet-SIINFEKL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7514)
  AUTHORS   Freed-Pastor WA, Lambert LJ, Ely ZA, Pattada NB, Bhutkar A, Eng G, 
            Mercer KL, Garcia AP, Lin L, Rideout WM 3rd, Hwang WL, Schenkel JM, 
            Jaeger AM, Bronson RT, Westcott PMK, Hether TD, Divakar P, Reeves 
            JW, Deshpande V, Delorey T, Phillips D, Yilmaz OH, Regev A, Jacks T
  TITLE     The CD155/TIGIT axis promotes and maintains immune evasion in 
            neoantigen-expressing pancreatic cancer.
  JOURNAL   Cancer Cell. 2021 Oct 11;39(10):1342-1360.e14. doi: 
            10.1016/j.ccell.2021.07.007. Epub 2021 Aug 5.
  PUBMED    34358448
REFERENCE   2  (bases 1 to 7514)
  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: 
            "10.1016/j.ccell.2021.07.007"; journalName: "Cancer Cell"; date: 
            "2021-10-11- 11"; volume: "39"; issue: "10"; pages: "1342-1360.e14"
COMMENT     Sequence Label: H11-mScarletSIIN
FEATURES             Location/Qualifiers
     source          1..7514
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(142..147,148..165,166..172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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)."
     primer_bind     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1181..1680
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1216..1239)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1613..1632
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1681..1690
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1692..1725)
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1737..1871
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1774..1793)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1828..1847
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(1900..1933)
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1946..1969
                     /codon_start=1
                     /product="antigenic CD8+ T cell epitope from chicken 
                     ovalbumin"
                     /label=OVA peptide
                     /translation="SIINFEKL"
     CDS             1991..2047
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2048..2740
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGGA
                     VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS
                     TGGMDELYK"
     primer_bind     complement(2178..2196)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2446..2465
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2779..3367
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2832..2852)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3250..3261)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3378..3394)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3407..3424)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3413..3637
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5118..5135)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5118..5134)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5127..5149)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5343..5362)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5462..5484
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5522..5540)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5608..5712
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5713..5781,5782..6573)
                     /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     complement(5931..5950)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6744..7332
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7233..7252
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7486..7503
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttac
      241 gtaaacagtc ttccagggga taagaacctt acagaaaaag cagaaatgcc tggggcactg
      301 gattaccgat gtaatcaaat tcagtccttg aattgaacac aggattgcct agagcaaggc
      361 cagccagaga ttcatctcag agggagaaag gtgtctttgg agcaattttg tggtaatcta
      421 gtatgtatca cataagttta gacgcatttg ggactggaaa gatgtgaaca aagcacccta
      481 tggctcacat ctgtcattaa ctctagttcc agtgcacctg acaccgtctt ctggcctctg
      541 cagtgaccaa gcacatgggt agtatgtaga catatacata agcaaaacac acatcattaa
      601 aaagtgacat ttcccaaagg aagctgaaga accagttctt gagaagatag tagaaatcag
      661 aaggggaaat agtagacata cagagggact gaccaggttg tgtcaccttt ataggctagg
      721 ctaatggatg atcgacacta gcgctctttg tgaaggacac acaaatgaga catagtttat
      781 aggactaaac acacttctaa gcaatttaat gagacttaag accctgtctc tagcaaatac
      841 tctggatgat attcagctca aggctcttgt cagacatgtt tccattttca aggtgagcta
      901 actggccaaa actgccaaca acctgtagtg aatagagaag atgagaaaat ctggattctc
      961 aaatgaccta atgaaagcca ctggagcgcc atatggtttc tgtgaaaatg ccttttcaat
     1021 cattaacctc ttaaatgagt gttagcatcc taactaatga gtggtgcaga atagtgggtc
     1081 tgcttagctt aactaaggcc aagaaaacaa aacaggaaat tcattccatg tcatgagact
     1141 catactgttt gatcagtgtg agggagtgta aagctggttt gggtagggga ggcgcttttc
     1201 ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt
     1261 ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt
     1321 ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg
     1381 cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtgtcactag tctcgtgcag
     1441 atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag
     1501 ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc
     1561 aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc
     1621 acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg
     1681 gccaccatgg ttaccgttcg tatagcatac attatacgaa gttatttgtt gttgttaact
     1741 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     1801 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     1861 atgtctggat ctgcgactct agaggatcat aatcagccat accgttcgta tagcatacat
     1921 tatacgaagt tatgagcagc tcgagagtat aatcaacttt gaaaaactgt cagcagaggc
     1981 tggcagcggc gccacgaact tctctctgtt aaagcaagca ggagacgtgg aagaaaaccc
     2041 cggtcctgtg agcaagggcg aggcagtgat caaggagttc atgcggttca aggtgcacat
     2101 ggagggctcc atgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     2161 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttctcctg
     2221 ggacatcctg tcccctcagt tcatgtacgg ctccagggcc ttcaccaagc accccgccga
     2281 catccccgac tactataagc agtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     2341 cttcgaggac ggcggcgccg tgaccgtgac ccaggacacc tccctggagg acggcaccct
     2401 gatctacaag gtgaagctcc gcggcaccaa cttccctcct gacggccccg taatgcagaa
     2461 gaagacaatg ggctgggaag cgtccaccga gcggttgtac cccgaggacg gcgtgctgaa
     2521 gggcgacatt aagatggccc tgcgcctgaa ggacggcggc cgctacctgg cggacttcaa
     2581 gaccacctac aaggccaaga agcccgtgca gatgcccggc gcctacaacg tcgaccgcaa
     2641 gttggacatc acctcccaca acgaggacta caccgtggtg gaacagtacg aacgctccga
     2701 gggccgccac tccaccggcg gcatggacga gctgtacaag taaaaacgtt gttgtttggg
     2761 gttgaattac tcttcggtaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     2821 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     2881 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     2941 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3001 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3061 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3121 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     3181 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     3241 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     3301 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     3361 tccccgcatc cccgggggat ccactagttc tagagcggcc gatcagcctc gactgtgcct
     3421 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3481 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3541 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3601 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3661 tggggctcga gatccactag ttctagcctc gaggctagag caaacacgag gttcccttag
     3721 acctcaggag aaaaagtctt tggctgtaag aacacacctc agtggatgtg gtagactatg
     3781 cctttactct tttttttttt tttttttttt tttgggtttt ttcgagacag ggtttctctg
     3841 tgtagccttg gctgtcctgg aactcacttt gttgaccagg ctggcctcga actcagagat
     3901 ccgcctgcct ctgcctctca agtgctagaa ttaaaagtgt gcgccaccac tcccagctat
     3961 atagactatg cctttaatca ggacacttgg gaggcagggg gatctcaaat tcaattccag
     4021 acagatgact gacaaacaca cacatacacc attcgtcttt tctttttttc ttttttcttt
     4081 tgaaaacagg tttctctatg cagccctggc tttcctagaa ttcaatctat agacccagct
     4141 ggcctcaaac acagaacctc ttgcctctgc ctcctcaaca ctatgactaa aggtgtgaac
     4201 caccaccacc accacccacc tggccaaaga atgaattgag tgaataagta cgagtgcaac
     4261 tctccagcag cctgaggcaa gaaagcttaa cagttgaacc tgagacccta cttcgtttgt
     4321 gttgtacata tcttctcata atgaactagg cagcctagtc tctctaccaa atacaccatg
     4381 cacttacacc tcctcaaacc aactgcaaca gctacacaca tttccatggc ttgcctgtaa
     4441 taataaattc acaaacttcc tctaagtaac ttttagtgga aatctaccag gttatgccta
     4501 tttgtgattg ggtgcacaga tgactagatg agatgtaatt tgggaagtga catgcaatag
     4561 gaaaaatgaa tatataacct ataaatagaa aacaccaagg caagcgtggt gactcacacc
     4621 tttaattagc acttgggagg cagaagtcgc atcatctctt aggtcaaggc caatctagtt
     4681 ccagaccagc aaggacaaca gaaaccttgc cttgatataa acattttaaa actttaaaaa
     4741 tagagaagcc caaactctgc tcattggtaa tctacccttc ctttctttag acaccttgag
     4801 aagctaggct acatagcttt gggtccttac gtacccccac tcactcagcc tgcagtcaat
     4861 ctctgatagt gtttcctgcc ataagctaca ctactgtttt gctccaaacc ttacttctag
     4921 tacaaatcca tgtgaggtct tcgtttcaat ttggggggga agaggccaga acacttgccg
     4981 cagatcccaa caacgggcag cacattccca gctagtgtat ctgatacaaa ctctatcttc
     5041 attcagaacc atcaagtcta agccccaaga gcaaaggtaa attcatgtat gtttggctgg
     5101 agaaagcatt tcaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     5161 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     5221 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg
     5281 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     5341 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     5401 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     5461 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     5521 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5581 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5641 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5701 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5761 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5821 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5881 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5941 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6001 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6061 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6121 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6181 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6241 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6301 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6361 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6421 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6481 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6541 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6601 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6661 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6721 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6781 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6841 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6901 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6961 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     7021 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7081 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7141 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7201 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7261 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7321 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7381 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7441 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     7501 gaggaagcgg aaga
//
