LOCUS       sequence-464240-        6496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expresses SP2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6496)
  AUTHORS   Chou WC, Chen WT, Kuo CT, Chang YM, Lu YS, Li CW, Hung MC, Shen CY
  TITLE     Genetic insights into carbohydrate sulfotransferase 8 and its impact
            on the immunotherapy efficacy of cancer.
  JOURNAL   Cell Rep. 2024 Jan 23;43(1):113641. doi: 
            10.1016/j.celrep.2023.113641. Epub 2023 Dec 30.
  PUBMED    38165805
REFERENCE   2  (bases 1 to 6496)
  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 Rep.";
            date: "2024-01-23"; volume: "43(1):113641. doi"; pages: " 
            10.1016/j.celrep.2023.113641. Epub 2023 Dec 30"
COMMENT     Sequence Label: pXJ-Myc-SP2
FEATURES             Location/Qualifiers
     source          1..6496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..305
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..509
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     460..480
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     506..530
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          671..1243
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     1251..1270
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     promoter        1298..1316
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1337..1366
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(3296..3314)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3588..3709
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3625..3644)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3679..3698
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3737..3754)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3908..4496)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3988..4007)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4667..5458,5459..5527))
                     /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     5290..5309
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5528..5632)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5700..5718
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5756..5778)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5878..5897
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5964..6419)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6101..6122)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6313..6332
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ccgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
       61 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      121 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      181 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      241 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      301 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      361 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      421 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      481 tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac
      541 gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccgcggcc
      601 gggaacggtg cattggaacg gacccctagg cttttgcaaa aagctccgga tcgatcctga
      661 gaacttcagg gtgagtttgg ggacccttga ttgttctttc tttttcgcta ttgtaaaatt
      721 catgttatat ggagggggca aagttttcag ggtgttgttt agaatgggaa gatgtccctt
      781 gtatcaccat ggaccctcat gataattttg tttctttcac tttctactct gttgacaacc
      841 attgtctcct cttattttct tttcattttc tgtaactttt tcgttaaact ttagcttgca
      901 tttgtaacga atttttaaat tcacttttgt ttatttgtca gattgtaagt actttctcta
      961 atcacttttt tttcaaggca atcagggtat attatattgt acttcagcac agttttagag
     1021 aacaattgtt ataattaaat gataaggtag aatatttctg catataaatt ctggctggcg
     1081 tggaaatatt cttattggta gaaacaacta catcctggtc atcatcctgc ctttctcttt
     1141 atggttacaa tgatatacac tgtttgagat gaggataaaa tactctgagt ccaaaccggg
     1201 cccctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct
     1261 ggttattgtg ctgtctcatc attttggcaa agaattgtaa tacgactcac tatagggcga
     1321 attcaccatg gcagaggagc aaaagctcat ttctgaagag gacttgttgc ggaaacatat
     1381 gggatccatg agcgatccac agaccagcat ggctgccact gctgctgtga gtcccagtga
     1441 ctacctgcag cctgccgcct ccaccaccca ggactcccag ccatctccct tagccctgct
     1501 tgctgcaaca tgtagcaaaa ttggccctcc agcagttgaa gctgctgtga cacctcctgc
     1561 tcccccacag cccacaccgc ggaaacttgt ccctatcaaa cctgcccctc tccctctcag
     1621 ccccggcaag aatagctttg gaatcttgtc ctccaaagga aatatacttc agattcaggg
     1681 gtcacaactg agcgcctcct atcctggagg gcagctggtg ttcgctatcc agaatcccac
     1741 catgatcaac aaagggaccc gatcaaatgc caatatccag taccaggcgg tccctcagat
     1801 tcaggcaagc aattcccaaa ccatccaagt acagcccaat ctcaccaacc agatccagat
     1861 catccctggc accaaccaag ccatcatcac cccctcaccg tccagtcaca agcctgtccc
     1921 catcaagcca gcccccatcc agaagtcgag tacgaccacc acccccgtgc agagcggggc
     1981 caatgtggtg aagctgacag gtgggggcgg caatgtgacg ctcactctgc ccgtcaacaa
     2041 ccttgtgaac gccagtgaca ccggggcccc tactcagctc ctcactgaaa gccccccaac
     2101 cccgctgtct aagactaaca agaaagcaag gaagaagagc cttcctgcct cccagccccc
     2161 tgtggctgtg gctgagcagg tggagacggt gctgatcgag accaccgcgg acaacatcat
     2221 ccaggcagga aataacctgc tcattgttca gagccctggt gggggccagc cagctgtggt
     2281 ccagcaggtc caggtggtgc cccccaaggc cgagcagcag caggtggtac agatccccca
     2341 gcaggctctg cgggtggtgc aggcggcatc tgccaccctc cccactgtac cccagaagcc
     2401 ctcccagaac tttcagatcc aggcagccga gccgacacct actcaggtct acatccgcac
     2461 gccttccggt gaggtgcaga cagtccttgt ccaggacagc cccccagcaa cagctgcagc
     2521 cacctctaac accacctgta gcagccctgc atcccgtgct ccccatctga gtgggaccag
     2581 caaaaagcac tcagctgcaa ttctccgaaa agagcgtccc ctgccaaaga ttgccccagc
     2641 cgggagcatc atcagcctga atgcagccca gttggcggca gctgcccagg caatgcagac
     2701 catcaacatc aatggtgtcc aggtccaggg cgtgcctgtc accatcacca acacaggcgg
     2761 gcagcagcag ctgacagtgc agaatgtttc tgggaacaac ctgaccatca gtgggctgag
     2821 ccccacccag atccagctgc aaatggaaca agccctggcc ggagagaccc agcccgggga
     2881 gaagcggcgc cgcatggcct gcacgtgtcc caactgcaag gatggggaga agaggtctgg
     2941 agagcagggc aagaagaagc acgtgtgcca catccccgac tgtggcaaga cgttccgtaa
     3001 gacgtccttg ctgcgtgccc atgtgcgcct gcacactggc gagcggccct ttgtctgcaa
     3061 ctggttcttc tgtgggaaga ggttcacacg gagtgacgag ctccaacggc atgctcgcac
     3121 ccacacaggg gacaaacgct tcgagtgcgc ccagtgtcag aagcgcttca tgaggagtga
     3181 ccacctcacc aagcattaca agacccacct ggtcacgaag aacttgtaag gccaactcga
     3241 gggggggccc ggtaccttaa ttaattaagg taccaggtaa gtgtacccaa ttcgccctat
     3301 agtgagtcgt attacaattc actcgatcgc ccttcccaac agttgcgcag cctgaatggc
     3361 gaatggagat ccaattttta agtgtataat gtgttaaact actgattcta attgtttgtg
     3421 tattttagat tcacagtccc aaggctcatt tcaggcccct cagtcctcac agtctgttca
     3481 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     3541 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     3601 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3661 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgcgactc
     3721 tagactcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3781 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3841 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3901 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3961 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4021 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4081 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4141 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4201 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4261 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4321 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4381 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4441 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4501 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4561 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4621 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4681 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4741 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4801 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4861 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4921 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4981 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5041 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5101 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5161 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5221 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5281 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5341 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5401 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5461 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5521 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5581 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5641 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     5701 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     5761 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     5821 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     5881 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     5941 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt
     6001 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     6061 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     6121 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     6181 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     6241 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     6301 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     6361 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     6421 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     6481 ttacgccagc tagagt
//
