LOCUS       sequence-113992-        6216 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of mouse Klf4..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6216)
  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 6216)
  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-Klf4
FEATURES             Location/Qualifiers
     source          1..6216
                     /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(3237..3256)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3302..3357
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3356..3375)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3718..3734)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3718..3734)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3731..3753)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3742..3758
                     /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(3766..3772,3773..3790,3791..3796))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3811..3832
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3891..4086
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3937..4072
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4092..4226
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4129..4148)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4183..4202
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4294..4311)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4465..5053)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4545..4564)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5224..6015,6016..6084))
                     /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     5847..5866
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6085..6189)
                     /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 tcatggctgt
     1741 cagcgacgct ctgctcccgt ccttctccac gttcgcgtcc ggcccggcgg gaagggagaa
     1801 gacactgcgt ccagcaggtg ccccgactaa ccgttggcgt gaggaactct ctcacatgaa
     1861 gcgacttccc ccacttcccg gccgccccta cgacctggcg gcgacggtgg ccacagacct
     1921 ggagagtggc ggagctggtg cagcttgcag cagtaacaac ccggccctcc tagcccggag
     1981 ggagaccgag gagttcaacg acctcctgga cctagacttt atcctttcca actcgctaac
     2041 ccaccaggaa tcggtggccg ccaccgtgac cacctcggcg tcagcttcat cctcgtcttc
     2101 cccagcgagc agcggccctg ccagcgcgcc ctccacctgc agcttcagct atccgatccg
     2161 ggccgggggt gacccgggcg tggctgccag caacacaggt ggagggctcc tctacagccg
     2221 agaatctgcg ccacctccca cggccccctt caacctggcg gacatcaatg acgtgagccc
     2281 ctcgggcggc ttcgtggctg agctcctgcg gccggagttg gacccagtat acattccgcc
     2341 acagcagcct cagccgccag gtggcgggct gatgggcaag tttgtgctga aggcgtctct
     2401 gaccacccct ggcagcgagt acagcagccc ttcggtcatc agtgttagca aaggaagccc
     2461 agacggcagc caccccgtgg tagtggcgcc ctacagcggt ggcccgccgc gcatgtgccc
     2521 caagattaag caagaggcgg tcccgtcctg cacggtcagc cggtccctag aggcccattt
     2581 gagcgctgga ccccagctca gcaacggcca ccggcccaac acacacgact tccccctggg
     2641 gcggcagctc cccaccagga ctacccctac actgagtccc gaggaactgc tgaacagcag
     2701 ggactgtcac cctggcctgc ctcttccccc aggattccat ccccatccgg ggcccaacta
     2761 ccctcctttc ctgccagacc agatgcagtc acaagtcccc tctctccatt atcaagagct
     2821 catgccaccg ggttcctgcc tgccagagga gcccaagcca aagaggggaa gaaggtcgtg
     2881 gccccggaaa agaacagcca cccacacttg tgactatgca ggctgtggca aaacctatac
     2941 caagagttct catctcaagg cacacctgcg aactcacaca ggcgagaaac cttaccactg
     3001 tgactgggac ggctgtgggt ggaaattcgc ccgctccgat gaactgacca ggcactaccg
     3061 caaacacaca gggcaccggc cctttcagtg ccagaagtgc gacagggcct tttccaggtc
     3121 ggaccacctt gccttacaca tgaagaggca ctttaattca ctcctcaggt gcaggctgcc
     3181 tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt
     3241 tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc
     3301 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
     3361 ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt
     3421 ttggcaacat atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat
     3481 cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga
     3541 ggttagattt tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt
     3601 tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct
     3661 gtccctcttc tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc
     3721 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3781 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3841 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat
     3901 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3961 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4021 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4081 tgcaaaaagc taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     4141 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     4201 tcaatgtatc ttatcatgtc tggatccgct gcattaatga atcggccaac gcgcggggag
     4261 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4321 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4381 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4441 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4501 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4561 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4621 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4681 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4741 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4801 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4861 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4921 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4981 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5041 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5101 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5161 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5221 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5281 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5341 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5401 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5461 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5521 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5581 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5641 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5701 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5761 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5821 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5881 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5941 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6001 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6061 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6121 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6181 ggttccgcgc acatttcccc gaaaagtgcc acctgg
//
