LOCUS       sequence-384387-        7222 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hipp11 targeting vector, expresses mScarlet-mALG8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7222)
  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 7222)
  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-mScarletALG8
FEATURES             Location/Qualifiers
     source          1..7222
                     /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(4826..4843)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4826..4842)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4835..4857)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5051..5070)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5170..5192
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5230..5248)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5316..5420
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5421..5489,5490..6281)
                     /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(5639..5658)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6452..7040
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6941..6960
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7194..7211
                     /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 agtgtgcagt gggcatcaca
     2401 tacacctgga ccaggctgta 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 tgggtttttt cgagacaggg
     3541 tttctctgtg tagccttggc tgtcctggaa ctcactttgt tgaccaggct ggcctcgaac
     3601 tcagagatcc gcctgcctct gcctctcaag tgctagaatt aaaagtgtgc gccaccactc
     3661 ccagctatat agactatgcc tttaatcagg acacttggga ggcaggggga tctcaaattc
     3721 aattccagac agatgactga caaacacaca catacaccat tcgtcttttc tttttttctt
     3781 ttttcttttg aaaacaggtt tctctatgca gccctggctt tcctagaatt caatctatag
     3841 acccagctgg cctcaaacac agaacctctt gcctctgcct cctcaacact atgactaaag
     3901 gtgtgaacca ccaccaccac cacccacctg gccaaagaat gaattgagtg aataagtacg
     3961 agtgcaactc tccagcagcc tgaggcaaga aagcttaaca gttgaacctg agaccctact
     4021 tcgtttgtgt tgtacatatc ttctcataat gaactaggca gcctagtctc tctaccaaat
     4081 acaccatgca cttacacctc ctcaaaccaa ctgcaacagc tacacacatt tccatggctt
     4141 gcctgtaata ataaattcac aaacttcctc taagtaactt ttagtggaaa tctaccaggt
     4201 tatgcctatt tgtgattggg tgcacagatg actagatgag atgtaatttg ggaagtgaca
     4261 tgcaatagga aaaatgaata tataacctat aaatagaaaa caccaaggca agcgtggtga
     4321 ctcacacctt taattagcac ttgggaggca gaagtcgcat catctcttag gtcaaggcca
     4381 atctagttcc agaccagcaa ggacaacaga aaccttgcct tgatataaac attttaaaac
     4441 tttaaaaata gagaagccca aactctgctc attggtaatc tacccttcct ttctttagac
     4501 accttgagaa gctaggctac atagctttgg gtccttacgt acccccactc actcagcctg
     4561 cagtcaatct ctgatagtgt ttcctgccat aagctacact actgttttgc tccaaacctt
     4621 acttctagta caaatccatg tgaggtcttc gtttcaattt gggggggaag aggccagaac
     4681 acttgccgca gatcccaaca acgggcagca cattcccagc tagtgtatct gatacaaact
     4741 ctatcttcat tcagaaccat caagtctaag ccccaagagc aaaggtaaat tcatgtatgt
     4801 ttggctggag aaagcatttc aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     4861 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     4921 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     4981 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
     5041 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     5101 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     5161 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     5221 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     5281 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     5341 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     5401 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     5461 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5521 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5581 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     5641 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5701 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     5761 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5821 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5881 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5941 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     6001 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     6061 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6121 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6181 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     6241 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     6301 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     6361 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6421 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6481 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6541 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6601 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6661 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6721 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6781 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6841 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6901 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6961 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     7021 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     7081 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     7141 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     7201 cagtgagcga ggaagcggaa ga
//
