LOCUS       sequence-113993-        5858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of mouse Klf2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5858)
  AUTHORS   Fiore C, Cohen BA
  TITLE     Interactions between pluripotency factors specify cis-regulation in 
            embryonic stem cells.
  JOURNAL   Genome Res. 2016 Jun;26(6):778-86. doi: 10.1101/gr.200733.115. Epub 
            2016 Apr 15.
  PUBMED    27197208
REFERENCE   2  (bases 1 to 5858)
  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: 
            "10.1101/gr.200733.115"; journalName: "Genome Res"; date: "2016-06";
            volume: "26"; issue: "6"; pages: "778-86"
COMMENT     Sequence Label: pCX-Klf2
FEATURES             Location/Qualifiers
     source          1..5858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..661
                     /label=chicken beta-actin promoter
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(2879..2898)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2944..2999
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2998..3017)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3360..3376)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3360..3376)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3373..3395)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3384..3400
                     /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(3408..3414,3415..3432,3433..3438))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3453..3474
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3533..3728
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3579..3714
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3734..3868
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3771..3790)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3825..3844
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3936..3953)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4107..4695)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4187..4206)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4866..5657,5658..5726))
                     /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     5489..5508
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5727..5831)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc
      541 ggggcgggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tatggcgctc
     1741 agcgagccta tcttgccgtc ctttgccact ttcgccagcc cgtgcgagcg cggcctccag
     1801 gagcgctggc cgcgaaatga acccgaggcg ggcggcacgg atgaggacct aaacaacgtg
     1861 ttggacttca tcctctccat gggattggac ggtctgggcg ccgagaatcc tcccgagccc
     1921 ccgccgcagc ccccgccgcc tgccttctac tacccggagc cgggtgctcc gccgccctac
     1981 agcatccccg cggccagcct gggaacagag ctgctgcgcc ccgacctgga cccgcctcag
     2041 gggccggctc tgcacggccg cttcctcctt gcgcctcccg ggcggctagt gaaggccgag
     2101 ccccccgagg tggacggcgg cggctacggc tgcgctccgg gcctggccca cggaccgcgc
     2161 gggctgaagc tcgagggcgc cccaggagcg acaggtgcat gcatgcgggg tcccgccggc
     2221 cgccccccgc cgcccccgga cacgccgccg ctcagccccg acggccccct gcgcatcccg
     2281 gcgtccggtc cacgcaaccc gttcccgccg cccttcggtc ccggccccag cttcggcggc
     2341 cccggccccg cgttgcacta cgggcctccc gcgcctggcg ccttcggtct tttcgaggac
     2401 gcggcggcag cggcggcggc gctgggcttg gctccacctg ccacgcgcgg tctcctcacg
     2461 ccgccctcgt ccccgctgga gctgctggag gccaagccca aacgcggccg ccgctcctgg
     2521 ccccgcaagc gcgccgccac acatacttgc agctacacca actgcggcaa gacctacacc
     2581 aagagctcgc acctaaaggc gcatctgcgt acacacacag gtgagaagcc ttatcattgc
     2641 aactgggaag gatgcggctg gaagttcgcg cgctctgacg agcttacccg ccactaccga
     2701 aagcacacag gtcacagacc ctttcagtgc cacttgtgcg accgggcctt ctccaggtcc
     2761 gaccacctgg ccttgcacat gaagcgacac atgtagaatt cactcctcag gtgcaggctg
     2821 cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc
     2881 tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg
     2941 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     3001 tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga
     3061 gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc
     3121 atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt
     3181 gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat
     3241 tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag
     3301 ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttggc gtaatcatgg
     3361 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3421 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     3481 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca
     3541 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3601 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3661 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3721 tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     3781 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3841 catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg
     3901 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3961 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4021 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4081 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4141 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4201 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4261 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4321 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4381 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4441 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4501 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4561 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4621 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4681 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4741 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4801 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4861 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4921 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4981 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5041 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5101 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5161 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5221 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5281 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5341 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5401 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5461 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5521 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5581 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5641 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5701 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5761 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5821 ggggttccgc gcacatttcc ccgaaaagtg ccacctgg
//
