LOCUS       sequence-384399-        7223 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hipp11 targeting vector, expresses mScarlet-mLAMA4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7223)
  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 7223)
  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-mScarletLAMA4
FEATURES             Location/Qualifiers
     source          1..7223
                     /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"
     CDS             1687..2382
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(1820..1838)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2088..2107
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2487..3075
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2540..2560)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2958..2969)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3086..3102)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3115..3132)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3121..3345
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4827..4844)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4827..4843)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4836..4858)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5052..5071)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5171..5193
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5231..5249)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5317..5421
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5422..5490,5491..6282)
                     /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(5640..5659)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6453..7041
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6942..6961
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7195..7212
                     /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 tgagcaaggg cgaggcagtg atcaaggagt tcatgcggtt caaggtgcac
     1741 atggagggct ccatgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     1801 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttctcc
     1861 tgggacatcc tgtcccctca gttcatgtac ggctccaggg ccttcaccaa gcaccccgcc
     1921 gacatccccg actactataa gcagtccttc cccgagggct tcaagtggga gcgcgtgatg
     1981 aacttcgagg acggcggcgc cgtgaccgtg acccaggaca cctccctgga ggacggcacc
     2041 ctgatctaca aggtgaagct ccgcggcacc aacttccctc ctgacggccc cgtaatgcag
     2101 aagaagacaa tgggctggga agcgtccacc gagcggttgt accccgagga cggcgtgctg
     2161 aagggcgaca ttaagatggc cctgcgcctg aaggacggcg gccgctacct ggcggacttc
     2221 aagaccacct acaaggccaa gaagcccgtg cagatgcccg gcgcctacaa cgtcgaccgc
     2281 aagttggaca tcacctccca caacgaggac tacaccgtgg tggaacagta cgaacgctcc
     2341 gagggccgcc actccaccgg cggcatggac gagctgtaca aggatggctt tgaagtaggc
     2401 ttcaatttcc gaacattata atgacgtacg gctaagcctc gactagggat aaaacgttgt
     2461 tgtttggggt tgaattactc ttcggtaatc aacctctgga ttacaaaatt tgtgaaagat
     2521 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     2581 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     2641 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     2701 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     2761 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     2821 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     2881 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     2941 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     3001 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     3061 gggccgcctc cccgcatccc cgggggatcc actagttcta gagcggccga tcagcctcga
     3121 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     3181 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     3241 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     3301 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     3361 gaaccagctg gggctcgaga tccactagtt ctagcctcga ggctagagca aacacgaggt
     3421 tcccttagac ctcaggagaa aaagtctttg gctgtaagaa cacacctcag tggatgtggt
     3481 agactatgcc tttactcttt tttttttttt tttttttttt ttgggttttt tcgagacagg
     3541 gtttctctgt gtagccttgg ctgtcctgga actcactttg ttgaccaggc tggcctcgaa
     3601 ctcagagatc cgcctgcctc tgcctctcaa gtgctagaat taaaagtgtg cgccaccact
     3661 cccagctata tagactatgc ctttaatcag gacacttggg aggcaggggg atctcaaatt
     3721 caattccaga cagatgactg acaaacacac acatacacca ttcgtctttt ctttttttct
     3781 tttttctttt gaaaacaggt ttctctatgc agccctggct ttcctagaat tcaatctata
     3841 gacccagctg gcctcaaaca cagaacctct tgcctctgcc tcctcaacac tatgactaaa
     3901 ggtgtgaacc accaccacca ccacccacct ggccaaagaa tgaattgagt gaataagtac
     3961 gagtgcaact ctccagcagc ctgaggcaag aaagcttaac agttgaacct gagaccctac
     4021 ttcgtttgtg ttgtacatat cttctcataa tgaactaggc agcctagtct ctctaccaaa
     4081 tacaccatgc acttacacct cctcaaacca actgcaacag ctacacacat ttccatggct
     4141 tgcctgtaat aataaattca caaacttcct ctaagtaact tttagtggaa atctaccagg
     4201 ttatgcctat ttgtgattgg gtgcacagat gactagatga gatgtaattt gggaagtgac
     4261 atgcaatagg aaaaatgaat atataaccta taaatagaaa acaccaaggc aagcgtggtg
     4321 actcacacct ttaattagca cttgggaggc agaagtcgca tcatctctta ggtcaaggcc
     4381 aatctagttc cagaccagca aggacaacag aaaccttgcc ttgatataaa cattttaaaa
     4441 ctttaaaaat agagaagccc aaactctgct cattggtaat ctacccttcc tttctttaga
     4501 caccttgaga agctaggcta catagctttg ggtccttacg tacccccact cactcagcct
     4561 gcagtcaatc tctgatagtg tttcctgcca taagctacac tactgttttg ctccaaacct
     4621 tacttctagt acaaatccat gtgaggtctt cgtttcaatt tgggggggaa gaggccagaa
     4681 cacttgccgc agatcccaac aacgggcagc acattcccag ctagtgtatc tgatacaaac
     4741 tctatcttca ttcagaacca tcaagtctaa gccccaagag caaaggtaaa ttcatgtatg
     4801 tttggctgga gaaagcattt caattcactg gccgtcgttt tacaacgtcg tgactgggaa
     4861 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     4921 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     4981 tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
     5041 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     5101 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     5161 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     5221 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     5281 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5341 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5401 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5461 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5521 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5581 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5641 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5701 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5761 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5821 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5881 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5941 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     6001 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6061 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6121 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6181 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6241 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6301 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6361 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6421 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6481 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6541 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6601 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6661 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6721 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6781 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6841 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6901 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6961 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7021 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7081 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     7141 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     7201 tcagtgagcg aggaagcgga aga
//
