LOCUS       sequence-464299-        5845 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expresses SP5 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5845)
  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 5845)
  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-SP5
FEATURES             Location/Qualifiers
     source          1..5845
                     /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(2645..2663)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2937..3058
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2974..2993)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3028..3047
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3086..3103)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3257..3845)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3337..3356)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4016..4807,4808..4876))
                     /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     4639..4658
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4877..4981)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5049..5067
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5105..5127)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5227..5246
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5313..5768)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5450..5471)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5662..5681
                     /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 gccgcggtgg ccgtcctccg gaacgactcg ctgcaggcct ttctccagga
     1441 ccgcaccccc agcgcctccc cggacctggg caagcactcg cccctggcat tgctggccgc
     1501 cacctgtagc cgcatcggcc agccgggcgc ggcggcgccc ccggacttcc tgcaggtgcc
     1561 ctacgacccc gcgctgggct caccctccag gctcttccac ccgtggacca ccgacatgcc
     1621 ggcgcactcg ccaggcgcac tgccgccccc gcatcccagc ttggggctga cgccgcagaa
     1681 gacgcacctg cagccgtcct tcggggctgc gcacgagctt ccccttacac cccccgccga
     1741 cccctcgtac ccctacgagt tctcgccggt caagatgctg ccctcgagca tggcggctct
     1801 gcccgccagc tgcgcgcccg cctacgtgcc ctacgcggcg caggccgcgc tgccgccagg
     1861 ctactccaac ctgctgcctc cgccgccgcc accgcccccg ccgcccacct gccgccagtt
     1921 gtcacccaac ccggcccccg acgacctccc gtggtggagc atcccgcagg cgggcgccgg
     1981 gccgggggcc tccggggttc cgggaagcgg cctctccggc gcctgtgccg gggcccccca
     2041 cgcgccccgc ttccccgcct ctgcggccgc tgctgctgcg gccgccgccg ccctacaaag
     2101 aggcctggtg ttgggcccgt cggactttgc gcagtaccag agccagatcg ccgcgctgct
     2161 gcagaccaag gcccccctgg cggccacggc caggaggtgc cgccgctgcc gctgtcccaa
     2221 ctgccaggcg gcgggcggcg cccccgaggc ggagccgggg aagaagaagc agcacgtgtg
     2281 ccacgtgccg ggctgcggca aggtgtacgg gaagacgtcg cacctgaagg cgcacctgcg
     2341 ctggcacacg ggcgagcgac ccttcgtgtg caactggctc ttctgcggga agagcttcac
     2401 gcgctcggac gagctgcagc ggcacctgcg gactcacacg ggcgagaagc gctttgcctg
     2461 tcccgagtgc ggcaagcgct tcatgcgcag cgaccacctc gcgaagcacg tcaagactca
     2521 ccagaataag aagctcaaag tcgctgaggc cggggttaag cgggaggacg cgcgggacct
     2581 gtgagtcgag ggggggcccg gtaccttaat taattaaggt accaggtaag tgtacccaat
     2641 tcgccctata gtgagtcgta ttacaattca ctcgatcgcc cttcccaaca gttgcgcagc
     2701 ctgaatggcg aatggagatc caatttttaa gtgtataatg tgttaaacta ctgattctaa
     2761 ttgtttgtgt attttagatt cacagtccca aggctcattt caggcccctc agtcctcaca
     2821 gtctgttcat gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     2881 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     2941 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3001 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac
     3061 gcgcgactct agactcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3121 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3181 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3241 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3301 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3361 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3421 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3481 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3541 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3601 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3661 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3721 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3781 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3841 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3901 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3961 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4021 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4081 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4141 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4201 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4261 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4321 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4381 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4441 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4501 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4561 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4621 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4681 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4741 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4801 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4861 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4921 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4981 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     5041 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     5101 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     5161 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     5221 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     5281 acagatgcgt aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa
     5341 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     5401 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     5461 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     5521 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     5581 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     5641 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     5701 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     5761 gggcgcgtcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     5821 tcttcgctat tacgccagct agagt
//
