LOCUS       sequence-467758-        7959 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Thy1.1 and Aoah overexpression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7959)
  AUTHORS   Gong L, Luo J, Yang E, Ru B, Qi Z, Yang Y, Rani A, Purohit A, Zhang 
            Y, Guan G, Paul R, Vu T, Chen Z, Ji R, Day CP, Wu C, Merlino G, 
            Fitzgerald D, Altan-Bonnet G, Aldape K, Wu J, Guan X, Jiang P
  TITLE     Cancer immunology data engine reveals secreted AOAH as a potential 
            immunotherapy.
  JOURNAL   Cell. 2025 Jul 24:S0092-8674(25)00793-7. doi: 
            10.1016/j.cell.2025.07.004.
  PUBMED    40730154
REFERENCE   2  (bases 1 to 7959)
  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"; journalName: "Cell. 2025 
            Jul 24:S0092-8674(25)00793-7. doi: 10.1016/j.cell.2025.07.004."
COMMENT     Sequence Label: MSCV-IRES-Thy1.1-Aoah
FEATURES             Location/Qualifiers
     source          1..7959
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     17..36
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(255..359)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     427..445
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(483..505)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     605..624
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     818..840
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     968..987
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             1187..1703
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    1767..2108
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             2175..2591
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2521..2543
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2559..2575
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    join(4345..4919,4920..4922,4923..4931)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4512..4529)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4739..4758
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4968..5456
                     /codon_start=1
                     /product="mouse GPI-anchored cell surface glycoprotein of 
                     the immunoglobulin superfamily"
                     /label=Thy1.1
                     /note="This allele of Thy1 is also known as CD90.1."
                     /translation="MNPAISVALLLSVLQVSRGQKVTSLTACLVNQNLRLDCRHENNTK
                     DNSIQHEFSLTREKRKHVLSGTLGIPEHTYRSRVTLSNQPYIKVLTLANFTTKDEGDYF
                     CELRVSGANPMSSNKSISVYRDKLVKCGGISLLVQNTSWMLLLLLSLSLLQALDFISL"
     LTR             5540..6054
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    6216..6232
                     /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(6240..6246,6247..6264,6265..6270))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6285..6306
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6423..6440)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6594..7182)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6674..6693)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7353..7959,1..185,186..254))
                     /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"
ORIGIN
        1 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
       61 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
      121 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
      181 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
      241 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
      301 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
      361 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
      421 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
      481 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
      541 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
      601 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
      661 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
      721 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
      781 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
      841 acggcgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc
      901 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt
      961 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
     1021 cggccacgat gcgtccggcg tagaggcgat tagtccaatt tgttaaagac aggatatcag
     1081 tggtccaggc tctagttttg actcaacaat atcaccagct gaagcctata gagtacgagc
     1141 catagataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca
     1201 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaatacata
     1261 actgagaata gagaagttca gatcaaggtt aggaacagag agacagcaga atatgggcca
     1321 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc
     1381 cagatgcggt cccgccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
     1441 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     1501 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc
     1561 gccagtcctc cgatagactg cgtcgcccgg gtacccgtat tcccaataaa gcctcttgct
     1621 gtttgcatcc gaatcgtgga ctcgctgatc cttgggaggg tctcctcaga ttgattgact
     1681 gcccacctcg ggggtctttc atttggaggt tccaccgaga tttggagacc cctgcccagg
     1741 gaccaccgac ccccccgccg ggaggtaagc tggccagcgg tcgtttcgtg tctgtctctg
     1801 tctttgtgcg tgtttgtgcc ggcatctaat gtttgcgcct gcgtctgtac tagttagcta
     1861 actagctctg tatctggcgg acccgtggtg gaactgacga gttctgaaca cccggccgca
     1921 accctgggag acgtcccagg gactttgggg gccgtttttg tggcccgacc tgaggaaggg
     1981 agtcgatgtg gaatccgacc ccgtcaggat atgtggttct ggtaggagac gagaacctaa
     2041 aacagttccc gcctccgtct gaatttttgc tttcggtttg gaaccgaagc cgcgcgtctt
     2101 gtctgctgca gcgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt
     2161 tgtctgaaaa ttagggccag actgttacca ctcccttaag tttgacctta ggtcactgga
     2221 aagatgtcga gcggatcgct cacaaccagt cggtagatgt caagaagaga cgttgggtta
     2281 ccttctgctc tgcagaatgg ccaaccttta acgtcggatg gccgcgagac ggcaccttta
     2341 accgagacct catcacccag gttaagatca aggtcttttc acctggcccg catggacacc
     2401 cagaccaggt cccctacatc gtgacctggg aagccttggc ttttgacccc cctccctggg
     2461 tcaagccctt tgtacaccct aagcctccgc ctcctcttcc tccatccgcc ccgtctctcc
     2521 cccttgaacc tcctcgttcg accccgcctc gatcctccct ttatccagcc ctcactcctt
     2581 ctctaggcgc cggaattcgt taacgccacc atgaagtttc cctggaaagt cttcaagacc
     2641 acccttctgc ttcttctgct ctctcactca ttggcctctg tgccctctga ggaccagcca
     2701 ggtgacagct actcccatgg ccaaagctgt ttagggtgtg tggtactggt atctgtgatt
     2761 gagcaactgg ctgaagttca caactcttca gtccaggtgg cgatggagag actctgcagc
     2821 tacctgcctg aaaagctgtt cttgaagacc gcctgctatt tcttggttca gacatttggc
     2881 tcagacatca taaaactact cgatgaagct atgaaggctg atgtggtgtg ctatgctctg
     2941 gaattttgca agcggggtgc tgtccaacca cagtgtcatc tctacccact tccccaggag
     3001 gcctgggagt ctgcactgga gaaggcgaga caagtcctca ggaggtcctc taccatgaaa
     3061 tatcccagaa gtggtcgtaa catttgttcc ctaccatttt tgaccaagat ctgccagaaa
     3121 attgaattat ctataaagaa ggccgtgcca ttcaaagaca tagattcaga caagcacagt
     3181 gttttcccaa cgctgcgggg ctaccactgg cgagggcgtg actgtaatga cagtgacaag
     3241 acagtgtacc cgggcagaag gccagacaac tgggatatcc atcaggactc caactgtaat
     3301 ggcatctggg gtattgatcc gaaagatgga attccatatg agaagaaatt ctgtgaaggt
     3361 tcacagccca ggggaatcat tttgctggga gactcagctg gggctcattt tcacatcccc
     3421 cctgaatggc tgacagcttc acagatgtct gtgaactcct tcctcaacct gccgtccgcc
     3481 cttaccgatg agctcaactg gccccaactc tctggtgtaa ctggatttct ggactccacg
     3541 agtggaattg aagaaaaatc catttaccat cgtttgagaa agagaaacca ttgcaatcac
     3601 agagactatc agagtatctc aaaaaatggt gcatcttcca gaaatctgaa gaattttatt
     3661 gaaagcttgt ctaggaacca ggcatcagac catccagcca ttgtccttta tgccatgatt
     3721 ggaaacgatg tttgtaattc gaaagctgac acagtaccag aaatgaccac tcctgaacaa
     3781 atgtacgcca acgtcatgca gactctgaca catctaaatt ctcacctgcc caatggcagc
     3841 cacgttattt tatacggctt accagatgga acttttctct gggatagctt gcacaaccga
     3901 taccaccctc taggccagct aaataaagat gtgacctatg cacagttctt ctcctttcta
     3961 agatgcctgc agctcaatcc ctgtaacggc tggatgtctt ccaacaagac cctccggact
     4021 ctcacttcag agagagccga acaactctcc aatacactga aaaaaattgc aaccaccgag
     4081 acgtttgcaa acttcgatct tttctacgtg gattttgcct tccatgaaat aatagaggac
     4141 tggcagaaaa gaggtggaca gccatggcag cttattgaac ctgtggatgg attccacccc
     4201 aatgaggtgg cttcacttct gcaagcaaat cgtgtctggg aaaagataca gcttcaatgg
     4261 cctcatgtcc tgggaaagga gaatccattc aactctcaga ttgaagaggt gtttggagac
     4321 caaggaggac actagccgcg gccgcccctc tccctccccc ccccctaacg ttactggccg
     4381 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc
     4441 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
     4501 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
     4561 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
     4621 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
     4681 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
     4741 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     4801 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa
     4861 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata
     4921 tggccacaac catgggtctt ttctgcagtc accgtcctcg aggcaccatg aacccagcca
     4981 tcagcgtcgc tctcctgctc tcagtcttgc aggtgtcccg agggcagaag gtgaccagcc
     5041 tgacagcctg cctggtgaac caaaaccttc gcctggactg ccgccatgag aataacacca
     5101 aggataactc catccagcat gagttcagcc tgacccgaga gaagaggaag cacgtgctct
     5161 caggcaccct tgggataccc gagcacacgt accgctcccg cgtcaccctc tccaaccagc
     5221 cctatatcaa ggtccttacc ctagccaact tcaccaccaa ggatgagggc gactactttt
     5281 gtgagcttcg cgtctcgggc gcgaatccca tgagctccaa taaaagtatc agtgtgtata
     5341 gagacaagct ggtcaagtgt ggcggcataa gcctgctggt tcagaacaca tcctggatgc
     5401 tgctgctgct gctttccctc tccctcctcc aagccctgga cttcatttct ctgtgacgcg
     5461 tagaattgga tccgtcgacc tgcagccaag cttatcgata aaataaaaga ttttatttag
     5521 tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt
     5581 aacgccattt tgcaaggcat ggaaaataca taactgagaa tagagaagtt cagatcaagg
     5641 ttaggaacag agagacagca gaatatgggc caaacaggat atctgtggta agcagttcct
     5701 gccccggctc agggccaaga acagatggtc cccagatgcg gtcccgccct cagcagtttc
     5761 tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat
     5821 ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc
     5881 aataaaagag cccacaaccc ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc
     5941 gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt
     6001 ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt tcatgggtaa
     6061 cagtttcttg aagttggaga acaacattct gagggtagga gtcgaatatt aagtaatcct
     6121 gactcaatta gccactgttt tgaatccaca tactccaata ctcctgaaat agttcattat
     6181 ggacagcgca gaaagagctg gggagaattg tgaaattgtt atccgctcac aattccacac
     6241 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     6301 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     6361 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     6421 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     6481 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     6541 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     6601 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     6661 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     6721 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6781 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6841 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6901 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6961 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     7021 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     7081 aaaagagttg gtagctcttg atccggcaaa aaaaccaccg ctggtagcgg tggttttttt
     7141 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     7201 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     7261 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     7321 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     7381 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     7441 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     7501 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7561 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7621 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     7681 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7741 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7801 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7861 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7921 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgc
//
