LOCUS       sequence-404878-        6451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For DdCBE editing using Ddd9 at JAK2 site in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6451)
  AUTHORS   Huang J, Lin Q, Fei H, He Z, Xu H, Li Y, Qu K, Han P, Gao Q, Li B, 
            Liu G, Zhang L, Hu J, Zhang R, Zuo E, Luo Y, Ran Y, Qiu JL, Zhao KT,
            Gao C
  TITLE     Discovery of deaminase functions by structure-based protein 
            clustering.
  JOURNAL   Cell. 2023 Jul 20;186(15):3182-3195.e14. doi: 
            10.1016/j.cell.2023.05.041. Epub 2023 Jun 27.
  PUBMED    37379837
REFERENCE   2  (bases 1 to 6451)
  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: 
            "10.1016/j.cell.2023.05.041"; journalName: "Cell"; date: 
            "2023-07-20- 20"; volume: "186"; issue: "15"; pages: "3182-3195.e14"
COMMENT     Sequence Label: pCMV-TALE-L-JAK2-Ddd9-N
FEATURES             Location/Qualifiers
     source          1..6451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        132..335
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     286..306
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     377..396
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        377..395
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             421..486
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             493..513
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3169..3417
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             3460..3480
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3506..3523)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3512..3736
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3807..3823)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3807..3823)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3820..3842)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3831..3847
                     /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(3855..3861,3862..3879,3880..3885))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3900..3921
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4038..4055)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4209..4797)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4289..4308)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4968..5759,5760..5828))
                     /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     5591..5610
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5829..5933)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6012..6031)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(6203..6451,1..131)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
       61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      361 agagatccgc ggccgctaat acgactcact atagggagac ccaagctggc tagcaccatg
      421 gactacaaag atcacgacgg cgactacaag gaccatgaca tcgactataa ggacgatgac
      481 gacaagatgg cccctaagaa gaaaagaaag gtgggcatcc acagaggggt acctatgatg
      541 gtggacctgc ggacactggg ctacagccag cagcaacagg agaagattaa gcctaaggtg
      601 cggagcaccg tggcccaaca ccacgaggcc cttgtgggcc acggcttcac ccacgcccac
      661 atcgtagctc tgtcacagca cccagctgcc ctgggcacag tggccgtgaa gtaccaggac
      721 atgatcgctg ctctgcctga agccacccac gaggccattg tgggcgttgg caagcagtgg
      781 tccggcgcca gagccctgga agccctgctg accgtagctg gagagctgag aggacctcct
      841 ctgcagctgg acaccggcca gctgctgaag atcgctaagc ggggcggagt gacagccgtg
      901 gaagccgtgc acgcctggcg gaacgctctg accggcgccc ctctgaacct gaccccagat
      961 caggtggtgg cgatcgcctc ccacgacgga ggcaaacagg ccctggagac cgtgcagaga
     1021 ctgctgcctg tgctgtgcca agaccacggc ctcacacccg agcaggtggt ggccatcgcc
     1081 agccacgacg gaggcaaaca ggccctggaa accgtgcagc ggctgctccc tgtgctgtgt
     1141 caggcccacg gcctgacccc agatcaggtg gttgccatcg cgtctaacat cggaggcaag
     1201 caggctctgg agacagttca gcgcttactg cctgtgctgt gccaggccca cggcctgacc
     1261 cccgctcagg ttgtcgccat cgcctccaac ggcggcggca agcaagccct ggagaccgtg
     1321 cagagactgc tgccggtgct gtgccaggat catggcctga cccccgatca ggtggtggcc
     1381 atcgccagca acggcggcgg caagcaggcc ctggagacgg tgcagagact gctgccagtg
     1441 ctgtgccagg accacggcct gacccctgaa caggtcgtgg ccatcgcctc caatggcggt
     1501 ggcaagcagg ccctcgaaac cgtgcagaga ctgcttcccg tgctgtgcca ggcccacggc
     1561 ctgacacctg accaagtggt cgctatcgcc agccacgatg gcggcaagca ggctctagaa
     1621 accgttcaga gactgctccc tgtgctgtgt caggcccacg gcctgacacc tgcccaggtg
     1681 gtggccatcg cctctaacgg cggcggcaag caagccctgg aaacagtgca gaggctgctg
     1741 cccgtactgt gccaggatca cggactcaca cctgatcagg tggtggccat cgccaacaac
     1801 aacggcggaa agcaggccct ggagacagtc caaagactgc tgccagtgtt gtgccaggac
     1861 cacgggctga cacctgagca ggtagtggcc atcgcctcta acggaggcgg taaacaggcc
     1921 ctggagacag tgcagagact gctgcccgtc ctgtgtcagg cccacggcct gacccctgac
     1981 caggtggtag ctatcgccag ccacgatggc ggcaaacaag ccctggaaac cgtccagaga
     2041 ctgctgcctg tgctgtgcca ggcccacggg ctaacccctg cccaagtggt agcaattgcc
     2101 agcaacatcg gcggcaagca agccctggag accgtgcagc ggctgctgcc cgtgctgtgc
     2161 caggaccacg gactgacccc cgaccaggtg gtagccatcg cgagcaatgg aggcggaaag
     2221 caggccctcg agaccgtgca gcggctgctg cccgtgctgt gtcaggatca cggacttaca
     2281 ccagagcagg tggtggctat tgctagccac gacggcggca aacaggccct ggagaccgtg
     2341 caaagactgc tacccgttct gtgtcaggcc cacggactga cccccgacca ggtggtggcc
     2401 atcgccaata acaacggcgg aaagcaggct ctggagacag tgcaaagact ccttcctgtg
     2461 ctgtgccaag ctcacggcct gacacctgcc caggtcgtgg cgatcgcctc caatggcggc
     2521 ggtaagcagg ctctggagac cgttcagcgg ttactgcccg tgctgtgtca ggatcatggc
     2581 ctgacccccg agcaggtggt ggccatcgcc agcaacatcg gcggcagacc cgctctcgaa
     2641 agcatcgtgg cacagctgag ccgtcctgat cctgccctag ccgccctgac caacgaccac
     2701 ctggtggccc tggcctgcct gggcgggaga cccgccctgg atgccgtgaa gaagggcctg
     2761 ccccacgccc ctgctctgat caagcggaca aatagaagaa tccccgagag aaccagccac
     2821 agagtggccg gatccgggaa gagcctgagt gagagccaag ccacactgtc cgtggcccag
     2881 aggctgctgg caaccatagg agaggaaggg aaaacagctg gtgtactgga gctggatgga
     2941 gaacttatcc ctctagtgag tgggaagtct tctctgccca actatgcagc cagcggccac
     3001 gtggagggcc aggctgccct catcatgaga gaccggggcg ccacttcagg tagattgctc
     3061 attgacaacc cttccggcat ctgtggctac tgcaagtccc aggtggctac tctgctccct
     3121 gaaaatgcca ccctgcaggt tggcacccca ctgggctccg gaggaagcac caacctgtct
     3181 gacatcatcg agaaggagac aggcaagcag ctggtcatcc aggagagcat cctgatgctg
     3241 cccgaagaag tcgaagaagt gatcggaaac aagcctgaga gcgatatcct ggtccatacc
     3301 gcctacgacg agagtaccga cgaaaatgtg atgctgctga catccgacgc cccagagtat
     3361 aagccctggg ctctggtcat ccaggattcc aacggagaga acaaaatcaa aatgctgtct
     3421 ggcggctcaa aaagaaccgc cgacggcagc gaattcgagc ccaagaagaa gaggaaagtc
     3481 taatgagttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3541 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3601 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3661 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3721 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctcgat accgtcgacc
     3781 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     3841 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagccta gggtgcctaa
     3901 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     3961 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4021 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4081 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4141 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4201 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4261 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4321 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4381 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4441 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4501 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4561 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4621 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4681 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4741 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4801 gatcctttga tcttttctac ggggtctgac actcagtgga acgaaaactc acgttaaggg
     4861 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4921 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4981 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5041 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5101 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5161 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5221 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5281 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5341 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5401 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5461 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5521 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5581 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5641 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5701 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5761 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5821 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5881 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5941 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc gatctcccga tcccctaggg
     6001 tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     6061 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     6121 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
     6181 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
     6241 acggggtcat tagttcatag cccatatatt gagttccgcg ttacataact tacggtaaat
     6301 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     6361 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     6421 actgcccact tggcagtaca tcaagtgtat c
//
