LOCUS       sequence-51320-e        6437 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of codon-optimized Artemis..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6437)
  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 6437)
  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.Artemis
FEATURES             Location/Qualifiers
     source          1..6437
                     /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(3017..3034)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3023..3247
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3293..3721
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3380..3399)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3590..3611
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3730..3750)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     3809..3828
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4113..4234
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4150..4169)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4204..4223
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4283..4299)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4283..4299)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4296..4318)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4307..4323
                     /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(4331..4337,4338..4355,4356..4361))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4376..4397
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4514..4531)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4685..5270)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4765..4784)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5441..6232,6233..6301))
                     /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     6064..6083
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6302..6406)
                     /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 atgtcctcat ttgaagggca gatggcagaa taccccacca ttagcattga tagatttgat
      961 agggaaaacc tcagggcacg ggcttatttc ctgagccact gccataagga ccacatgaaa
     1021 gggctcaggg cacctaccct caagaggaga ctggagtgct ccctcaaagt ctacctgtat
     1081 tgttctccag tgacaaagga gctgctcctg acttccccca aatatcgctt ttggaagaaa
     1141 cgaatcattt ctatcgagat tgaaactcca acccagatca gtctggtgga tgaggcttca
     1201 ggcgaaaagg aggaaattgt ggtcaccctc ctgccagcag gacactgtcc aggtagcgtc
     1261 atgttcctgt ttcagggcaa caatggaacc gtgctgtaca caggcgactt ccgcctcgct
     1321 cagggagagg cagctcgaat ggaactcctg cattctggcg gacgggtcaa ggatatccag
     1381 agtgtgtatc tggacaccac attctgcgat ccccggtttt accagattcc tagccgcgag
     1441 gaatgtctgt ccggagtgct ggagctggtg aggtcatgga tcaccagaag cccatatcac
     1501 gtggtctggc tgaactgcaa ggcagcctac gggtatgagt acctcttcac aaatctgtcc
     1561 gaggaactcg gtgtgcaggt ccatgtgaac aaactggaca tgtttcgcaa tatgcccgag
     1621 atcctccacc atctgactac cgataggaac acccagattc acgcttgcag acatcccaag
     1681 gcagaggaat acttccagtg gagtaaactg ccttgtggca tcacttcacg gaaccgcatt
     1741 cccctccaca tcattagcat caagccttcc accatgtggt ttggcgagcg atccaggaaa
     1801 accaatgtca ttgtgcgaac aggagaaagc tcctataggg cctgcttctc ttttcattct
     1861 agttacagtg agatcaagga cttcctctct tatctgtgtc ctgtgaacgc ttaccctaat
     1921 gtcatcccag tgggcacaac tatggataag gtggtcgaga ttctcaaacc actgtgtcgg
     1981 tcaagccaga gcacagaacc caagtacaaa cctctcggaa agctgaaaag agcccggact
     2041 gtgcaccgag acagcgagga agaggacgat tatctgtttg acgatcccct gcctatccca
     2101 ctcagacaca aggtgcccta ccctgagact ttccatcccg aagtcttttc catgaccgct
     2161 gtgtctgaga agcagccaga aaaactgaga cagaccccag gatgctgtcg agcagagtgc
     2221 atgcagtcct ctaggttcac aaactttgtg gactgtgaag agtccaattc tgagagtgaa
     2281 gaggaagtgg gcatccccgc ctcactgcag ggggatctcg gtagcgtgct ccacctgcag
     2341 aaggctgacg gcgacgtccc acagtgggag gtgttcttta aaagaaacga cgaaatcacc
     2401 gatgagtccc tggaaaattt ccctagttca acagtggccg ggggttcaca gagcccaaag
     2461 ctgttttccg actctgatgg ggagtctact cacatcagct cccagaactc tagtcagagc
     2521 acacatatta ctgagcaggg ctcccaggga tgggacagtc agtcagatac agtcctggtg
     2581 tcaagccagg agcggaacag tggtgacatc acatcactgg acaaggcaga ttatcgccct
     2641 actatcaaag agaacattcc agccagcctg atggaacaga atgtgatttg ccctaaggac
     2701 acctactctg atctgaagag tagagacaaa gatgtcacta tcgtgcctag caccggcgag
     2761 ccaaccacac tgtcctctga aactcacatt cccgaggaaa agagcctcct gaacctgtcc
     2821 accaatgcag actctcagag ttcaagcgat ttcgaggtgc catctacacc cgaggccgaa
     2881 ctgcctaagc gggaacatct ccagtatctg tacgagaaac tggccacagg agaaagcatc
     2941 gctgtgaaga aacgcaagtg tagcctcctg gacactctcg agtaatctag agggcccgtt
     3001 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     3061 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     3121 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     3181 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     3241 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     3301 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3361 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3421 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3481 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3541 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3601 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3661 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3721 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3781 gaagtatgca aagcctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     3841 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     3901 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     3961 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4021 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4081 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     4141 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4201 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     4261 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     4321 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     4381 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4441 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4501 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4561 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4621 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4681 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4741 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4801 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4861 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4921 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4981 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5041 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5101 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5161 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5221 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5281 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5341 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5401 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5461 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5521 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5581 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5641 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5701 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5761 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5821 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5881 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5941 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6001 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6061 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6121 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6181 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6241 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6301 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6361 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6421 aagtgccacc tgacgtc
//
