LOCUS       sequence-51318-e        5100 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of codon-optimized Trex2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5100)
  AUTHORS   Certo MT, Gwiazda KS, Kuhar R, Sather B, Curinga G, Mandt T, Brault 
            M, Lambert AR, Baxter SK, Jacoby K, Ryu BY, Kiem HP, Gouble A, 
            Paques F, Rawlings DJ, Scharenberg AM
  TITLE     Coupling endonucleases with DNA end-processing enzymes to drive gene
            disruption.
  JOURNAL   Nat Methods. 2012 Oct;9(10):973-5. doi: 10.1038/nmeth.2177. Epub 
            2012 Sep 2.
  PUBMED    22941364
REFERENCE   2  (bases 1 to 5100)
  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: 
            "10.1038/nmeth.2177"; journalName: "Nat Methods"; date: "2012-10"; 
            volume: "9"; issue: "10"; pages: "973-5"
COMMENT     Sequence Label: pExodus CMV.Trex2
FEATURES             Location/Qualifiers
     source          1..5100
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1680..1697)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1686..1910
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1956..2384
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2043..2062)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2253..2274
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2393..2413)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     2472..2491
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2776..2897
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2813..2832)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2867..2886
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2946..2962)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2946..2962)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2959..2981)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2970..2986
                     /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(2994..3000,3001..3018,3019..3024))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3039..3060
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3177..3194)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3348..3933)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3428..3447)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4104..4895,4896..4964))
                     /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     4727..4746
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4965..5069)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 atgtctgagc cacctcgggc tgagaccttt gtattcctgg acctagaagc cactgggctc
      961 ccaaacatgg accctgagat tgcagagata tccctttttg ctgttcaccg ctcttccctg
     1021 gagaacccag aacgggatga ttctggttcc ttggtgctgc cccgtgttct ggacaagctc
     1081 acactgtgca tgtgcccgga gcgccccttt actgccaagg ccagtgagat tactggtttg
     1141 agcagcgaaa gcctgatgca ctgcgggaag gctggtttca atggcgctgt ggtaaggaca
     1201 ctgcagggct tcctaagccg ccaggagggc cccatctgcc ttgtggccca caatggcttc
     1261 gattatgact tcccactgct gtgcacggag ctacaacgtc tgggtgccca tctgccccaa
     1321 gacactgtct gcctggacac actgcctgca ttgcggggcc tggaccgtgc tcacagccac
     1381 ggcaccaggg ctcaaggccg caaaagctac agcctggcca gtctcttcca ccgctacttc
     1441 caggctgaac ccagtgctgc ccattcagca gaaggtgatg tgcacaccct gcttctgatc
     1501 ttcctgcatc gtgctcctga gctgctcgcc tgggcagatg agcaggcccg cagctgggct
     1561 catattgagc ccatgtacgt gccacctgat ggtccaagcc tcgaagcctg aattctgcag
     1621 atatccagca cagtggcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc
     1681 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     1741 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     1801 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     1861 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     1921 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     1981 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     2041 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2101 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2161 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     2221 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     2281 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     2341 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     2401 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagccta
     2461 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     2521 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     2581 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     2641 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     2701 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     2761 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     2821 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     2881 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     2941 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     3001 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     3061 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3121 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3181 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3241 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3301 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3361 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3421 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3481 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3541 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3601 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3661 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3721 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3781 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3841 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag
     3901 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3961 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4021 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4081 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4141 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4201 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4261 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4321 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4381 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4441 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4501 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4561 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4621 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4681 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4741 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4801 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4861 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4921 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4981 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5041 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
