LOCUS       sequence-464238-        5716 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expresses KLF2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5716)
  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 5716)
  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-KLF2
FEATURES             Location/Qualifiers
     source          1..5716
                     /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(2516..2534)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2808..2929
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2845..2864)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2899..2918
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2957..2974)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3128..3716)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3208..3227)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3887..4678,4679..4747))
                     /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     4510..4529
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4748..4852)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4920..4938
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4976..4998)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5098..5117
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5184..5639)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5321..5342)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5533..5552
                     /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 gcgctgagtg aacccatcct gccgtccttc tccactttcg ccagcccgtg
     1441 ccgcgagcgc ggcctgcagg agcgctggcc gcgcgccgaa cccgagtccg gcggcaccga
     1501 cgacgacctc aacagcgtgc tggacttcat cctgtccatg gggctggatg gcctgggcgc
     1561 cgaggccgcc ccggagccgc cgccgccgcc cccgccgcct gcgttctatt accccgaacc
     1621 cggcgcgccc ccgccctaca gcgcccccgc gggtggcctg gtgtctgagc tgctgcgacc
     1681 cgagctggat gcgccgctgg ggcccgcact gcacggccgc tttctgctgg cgccgcccgg
     1741 ccgcctggtc aaggccgagc cccctgaagc ggacggcggc ggcggctacg gctgcgcccc
     1801 cgggctgacc cgtggaccgc gcggcctcaa gcgcgagggc gccccgggcc cggcggcttc
     1861 gtgcatgcga ggtcccgggg gccgcccccc gccgccgccc gacacaccgc cgctcagccc
     1921 cgacggcccc gcgcgcctgc ccgcgcccgg tccgcgcgcc tccttcccgc cgcctttcgg
     1981 tggccctggt ttcggcgcgc ccgggcccgg cctgcattac gcgccgcctg cgcccccagc
     2041 cttcggtctc ttcgacgacg cggccgccgc cgcggcagcc ctgggcctgg cgccccccgc
     2101 cgcccgcggt ctcctcacgc cgcctgcgtc cccgctggag ctgctggagg ccaagccaaa
     2161 gcgcggccgc cgctcttggc cccgcaaacg caccgccact cacacctgca gctacgcggg
     2221 ctgcggcaag acctacacca agagttcgca tctgaaggcg catctgcgca cgcacacagg
     2281 tgagaagccc taccactgca actgggacgg ctgcggctgg aagtttgcgc gctcagacga
     2341 gctcacgcgc cactaccgaa agcacacggg ccaccggcca ttccagtgcc atctgtgcga
     2401 tcgtgccttc tcgcgctccg atcacctggc gctgcacatg aaacggcaca tgtagctcga
     2461 gggggggccc ggtaccttaa ttaattaagg taccaggtaa gtgtacccaa ttcgccctat
     2521 agtgagtcgt attacaattc actcgatcgc ccttcccaac agttgcgcag cctgaatggc
     2581 gaatggagat ccaattttta agtgtataat gtgttaaact actgattcta attgtttgtg
     2641 tattttagat tcacagtccc aaggctcatt tcaggcccct cagtcctcac agtctgttca
     2701 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     2761 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     2821 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2881 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgcgactc
     2941 tagactcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3001 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3061 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3121 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3181 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3241 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3301 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3361 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3421 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3481 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3541 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3601 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3661 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3721 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3781 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3841 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3901 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3961 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4021 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4081 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4141 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4201 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4261 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4321 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4381 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4441 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4501 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4561 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4621 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4681 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4741 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4801 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4861 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     4921 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     4981 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     5041 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     5101 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     5161 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt
     5221 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     5281 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     5341 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     5401 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     5461 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     5521 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     5581 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     5641 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     5701 ttacgccagc tagagt
//
