LOCUS       sequence-175032-        5557 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mouse Trex2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5557)
  AUTHORS   Yumlu S, Bashir S, Stumm J, Kuhn R
  TITLE     Efficient Gene Editing of Human Induced Pluripotent Stem Cells Using
            CRISPR/Cas9.
  JOURNAL   Methods Mol Biol. 2019;1961:137-151. doi: 
            10.1007/978-1-4939-9170-9_10.
  PUBMED    30912045
REFERENCE   2  (bases 1 to 5557)
  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: "Methods Mol
            Biol."; date: "2019"; volume: "1961:137-151. doi"; pages: " 
            10.1007/978-1-4939-9170-9_10"
COMMENT     Sequence Label: pCAG-Trex2-bpA
FEATURES             Location/Qualifiers
     source          1..5557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        15..394
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        396..673
                     /label=chicken beta-actin promoter
     intron          674..1685
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1609..1631
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1698..1717
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1698..1716
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2455..2471)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2485..2502)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2491..2715
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2787..2804)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2787..2803)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2796..2818)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2944..3399)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3081..3102)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3293..3312
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3426..3530
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3531..3599,3600..4391)
                     /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     complement(3749..3768)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4562..5150
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5051..5070
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5304..5321
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5438..5459
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5474..5479,5480..5497,5498..5504)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5512..5528
                     /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)."
     primer_bind     5517..5539
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5536..5552
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5536..5552
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ggcgcgccgg attcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
       61 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      121 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      181 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      241 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      301 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      361 gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt
      421 cactctcccc atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt
      481 attttgtgca gcgatggggg cggggggggg gggggggcgc gcgccaggcg gggcggggcg
      541 gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc
      601 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
      661 cgcgcggcgg gcgggagtcg ctgcgtcgcg ccttcgcccc gtgccccgct ccgccgccgc
      721 ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga
      781 cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggctcg tttcttttct
      841 gtggctgcgt gaaagcctta aagggctccg ggagggccct ttgtgcgggg gggagcggct
      901 cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggcccg cgctgcccgg
      961 cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg ctccgcgtgt gcgcgagggg
     1021 agcgcggccg ggggcggtgc cccgcggtgc gggggggctg cgaggggaac aaaggctgcg
     1081 tgcggggtgt gtgcgtgggg gggtgagcag ggggtgtggg cgcggcggtc gggctgtaac
     1141 ccccccctgc acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc
     1201 cgtgcggggc gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg
     1261 ccgggcgggg cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc
     1321 ggagcgccgg cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt
     1381 gcgagagggc gcagggactt cctttgtccc aaatctggcg gagccgaaat ctgggaggcg
     1441 ccgccgcacc ccctctagcg ggcgcgggcg aagcggtgcg gcgccggcag gaaggaaatg
     1501 ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc ttctccatct ccagcctcgg
     1561 ggctgccgca gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct
     1621 ggcgtgtgac cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc
     1681 tacagatcct taattaataa tacgactcac tataggggcc gccaccatgt ctgagccacc
     1741 ccgggctgag acctttgtat tcctggacct agaagccact gggctcccaa acatggatcc
     1801 tgagattgca gagatatccc tttttgctgt tcaccgctct tccctggaga acccagaacg
     1861 ggatgattct ggttccttgg tgctgccccg tgttctggac aagctcacac tgtgcatgtg
     1921 cccggagcgc ccctttactg ccaaggccag tgagattact ggtttgagca gcgaaagcct
     1981 gatgcactgc gggaaggctg gtttcaatgg cgctgtggta aggacactgc agggcttcct
     2041 aagccgccag gagggcccca tctgccttgt ggcccacaat ggcttcgatt atgacttccc
     2101 actgctgtgc acggagctac aacgtctggg tgcccatctg ccccaagaca ctgtctgcct
     2161 ggacacactg cctgcattgc ggggcctgga ccgtgctcac agccacggca ccagggctca
     2221 aggccgcaaa agctacagcc tggccagtct cttccaccgc tacttccagg ctgaacccag
     2281 tgctgcccat tcagcagaag gtgatgtgca caccctgctt ctgatcttcc tgcatcgtgc
     2341 tcctgagctg ctcgcctggg cagatgagca ggcccgcagc tgggctcata ttgagcccat
     2401 gtacgtgcca cctgatggtc caagcctcga agcctgaacg cgtaaatgat tgcagatcca
     2461 ctagttctag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     2521 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     2581 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2641 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     2701 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctcgaga tccactagtt
     2761 ctagcctcga ggctagagcg gccgccactg gccgtcgttt tacaacgtcg tgactgggaa
     2821 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     2881 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     2941 tggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3001 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3061 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     3121 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     3181 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     3241 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     3301 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     3361 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     3421 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3481 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     3541 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     3601 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     3661 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     3721 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     3781 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     3841 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3901 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     3961 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4021 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     4081 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4141 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4201 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     4261 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4321 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     4381 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     4441 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4501 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4561 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4621 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4681 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4741 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4801 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4861 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4921 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4981 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5041 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5101 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5161 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     5221 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     5281 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     5341 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     5401 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     5461 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     5521 gataacaatt tcacacagga aacagctatg accatga
//
