LOCUS       sequence-245950-        7056 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dTdT in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7056)
  AUTHORS   Loveless TB, Grotts JH, Schechter MW, Forouzmand E, Carlson CK, 
            Agahi BS, Liang G, Ficht M, Liu B, Xie X, Liu CC
  TITLE     Lineage tracing and analog recording in mammalian cells by 
            single-site DNA writing.
  JOURNAL   Nat Chem Biol. 2021 Jun;17(6):739-747. doi: 
            10.1038/s41589-021-00769-8. Epub 2021 Mar 22.
  PUBMED    33753928
REFERENCE   2  (bases 1 to 7056)
  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/s41589-021-00769-8"; journalName: "Nat Chem Biol"; date: 
            "2021-06"; volume: "17"; issue: "6"; pages: "739-747"
COMMENT     Sequence Label: pJHG004
FEATURES             Location/Qualifiers
     source          1..7056
                     /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"
     regulatory      911..920
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2491..2508)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2497..2721
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2767..3195
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2854..2873)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3064..3085
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3204..3224)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3209..3538
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3389..3524
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3451..3470
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    4729..4850
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4766..4785)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4820..4839
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4899..4915)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4899..4915)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4912..4934)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4923..4939
                     /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(4947..4953,4954..4971,4972..4977))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4992..5013
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5130..5147)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5301..5889)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5381..5400)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6060..6851,6852..6920))
                     /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     6683..6702
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6921..7025)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cctgtgtgtt 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 gctaccggtc gccaccatgg atccaccacg agcgtcccac ttgagccctc ggaagaagag
      961 accccggcag acgggtgcct tgatggcctc ctctcctcaa gacatcaaat ttcaagattt
     1021 ggtcgtcttc attttggaga agaaaatggg aaccacccgc agagcgttcc tcatggagct
     1081 ggcccgcagg aaagggttca gggttgaaaa tgagctcagt gattctgtca cccacattgt
     1141 agcagagaac aactcgggtt cggatgttct ggagtggctt caagcacaga aagtacaagt
     1201 cagctcacaa ccagagctcc tcgatgtctc ctggctgatc gaatgcatag gagcagggaa
     1261 accggtggaa atgacaggaa aacaccagct tgttgtgaga agagactatt cagatagcac
     1321 caacccaggc cccccgaaga ctccaccaat tgctgtacaa aagatctccc agtatgcgtg
     1381 tcagagaaga accactttaa acaactgtaa ccagatattc acggatgcct ttgatatact
     1441 ggctgaaaac tgtgagttta gagaaaatga agactcctgt gtgacattta tgagagcagc
     1501 ttctgtattg aaatctctgc cattcacaat catcagtatg aaggacacag aaggaattcc
     1561 ctgcctgggg tccaaggtga agggtatcat agaggagatt attgaagatg gagaaagttc
     1621 tgaagttaaa gctgtgttaa atgatgaacg atatcaatcc ttcaaactct ttacttctgt
     1681 atttggagtg gggctgaaga cttctgagaa gtggttcagg atgggtttca gaactctgag
     1741 taaagtaagg tcggacaaaa gcctgaaatt tacacgaatg cagaaagcag gatttctgta
     1801 ttatgaagac cttgtcagct gtgtgaccag ggcagaagca gaggccgtca gtgtgctggt
     1861 taaagaggct gtctgggcat ttcttccgga tgctttcgtc accatgacag gagggttccg
     1921 gaggggtaag aagatggggc atgaggtaga gtttttaatt accagcccag gatcaacaga
     1981 ggatgaagag caacttttac agaaagtgat gaacttatgg gaaaagaagg gattactttt
     2041 atattatgac cttgtggagt caacatttga aaagctcagg ttgcctagca ggaaggttga
     2101 tgctttggat cattttcaaa agtgctttct gattttcaaa ttgcctcgtc aaagagtgga
     2161 cagtgaccag tccagctggc aggaaggaaa gacctggaag gccatccgtg tggatttagt
     2221 tctgtgcccc tacgagcgtc gtgcctttgc cctgttggga tggactggct cccggcagtt
     2281 tgagagagac ctccggcgct atgccacaca tgagcggaag atgattctgg ataaccatgc
     2341 tttatatgac aagaccaaga ggatattcct caaagcagaa agtgaagaag aaatttttgc
     2401 gcatctggga ttggattata ttgaaccgtg ggaaagaaat gcctaggcgg ccgctcgagt
     2461 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     2521 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     2581 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     2641 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     2701 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     2761 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2821 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     2881 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2941 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3001 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3061 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3121 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3181 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3241 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3301 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3361 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3421 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     3481 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     3541 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     3601 tcgcatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg
     3661 tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta
     3721 tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaatttag
     3781 cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc
     3841 tgaaaccgaa ctgcccgctg ttctgcaacc ggtcgcggag gccatggatg cgatcgctgc
     3901 ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata
     3961 cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac
     4021 tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg
     4081 ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt
     4141 cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga
     4201 ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca
     4261 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc
     4321 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga
     4381 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt
     4441 cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt
     4501 actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aataggcggg
     4561 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4621 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4681 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     4741 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4801 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     4861 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     4921 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4981 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     5041 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5101 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5161 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5221 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5281 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5341 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5401 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5461 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5521 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5581 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5641 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5701 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5761 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5821 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5881 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5941 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6001 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6061 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6121 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6181 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6241 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6301 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6361 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6421 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6481 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6541 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6601 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6661 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6721 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6781 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6841 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6901 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6961 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7021 ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
