LOCUS       sequence-415815-        5196 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Unfused B103 DNA polymerase, expressed from CMV or T7 promoters. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5196)
  AUTHORS   Ferreira da Silva J, Tou CJ, King EM, Eller ML, Rufino-Ramos D, Ma 
            L, Cromwell CR, Metovic J, Benning FMC, Chao LH, Eichler FS, 
            Kleinstiver BP
  TITLE     Click editing enables programmable genome writing using DNA 
            polymerases and HUH endonucleases.
  JOURNAL   Nat Biotechnol. 2024 Jul 22. doi: 10.1038/s41587-024-02324-x.
  PUBMED    39039307
REFERENCE   2  (bases 1 to 5196)
  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: "Nat 
            Biotechnol. 2024 Jul 22. doi: 10.1038/s41587-024-02324-x."
COMMENT     Sequence Label: Polymerase expression - pCMV-B103 (LM2907)
FEATURES             Location/Qualifiers
     source          1..5196
                     /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             2182..2202
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2211..2228
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2251..2268)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2257..2481
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2552..2568)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2552..2568)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2565..2587)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2576..2592
                     /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(2600..2606,2607..2624,2625..2630))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2645..2666
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2783..2800)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2954..3542)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3034..3053)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3713..4504,4505..4573))
                     /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     4336..4355
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4574..4678)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4757..4776)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(4948..5196,1..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 atagggagag ccgccaccat gcctagaaag
      421 atgttcagct gcgatttcga aaccacaacc aagctggacg actgcagagt gtgggcttac
      481 ggctacatgg aaatcggcaa cctggataac tacaagattg gaaacagcct ggatgagttc
      541 atgcagtggg tgatggaaat ccaggccgac ctgtacttcc acaacctgaa gtttgacggc
      601 gccttcatag tgaactggct ggaaagacac ggcttcaagt ggagcaacga gggcctgccc
      661 aacacctaca ataccatcat ttctaagatg ggccagtggt acatgatcga tatctgtttc
      721 ggctataagg gcaagcggaa gctgcacacc gtgatctacg acagcctgaa gaagctgcca
      781 ttccctgtca aaaagatcgc caaagatttt cagctgcctc tgctgaaagg cgatatcgac
      841 taccacaagg agcggcctgt cggccacgaa atcacaccag aggaatatga atacatcaag
      901 aatgacatcg agatcatcgc ccgggccctg gacatccagt tcaagcaagg cctggataga
      961 atgaccgctg gctctgattc tctgaagggg ttcaaagata tcctgagcac caagaagttc
     1021 aacaaggtgt tccccaagtt aagcctgcct atggacaagg aaatccgcta cgcctatagg
     1081 ggcggcttta catggctgaa cgacaaatac aaggaaaagg agattggaga aggcatggtg
     1141 ttcgatgtga actctctgta tccttcccag atgtacagca gacctctgcc ttacggcgcc
     1201 cccatcgtgt tccagggcaa gtacgagaag gacgagcagt accccctgta catccaaaga
     1261 atcagattcg agttcgagct gaaagagggc tatatcccca ccatccagat caaaaaaaac
     1321 cccttcttca agggcaatga gtacctgaag aactccggcg gagagcctgt tgaactgtac
     1381 ctgaccaacg tggatctgga actgatccag gagcactacg agctgtacaa cgtggagtac
     1441 attgacggct ttaaatttag agaaaaaacc ggcttgttta aggacttcat cgacaagtgg
     1501 acctacgtga agacacacga ggaaggagca aagaaacagc tggccaagct gatgctgaac
     1561 agcctgtacg gcaagtttgc cagcaatcct gacgtgaccg gcaaagttcc atacctcaaa
     1621 gaggacggat ctctgggctt ccgggtgggc gacgaagaat acaaggaccc cgtgtacacc
     1681 cctatgggag tgtttatcac agcctgggcc agattcacca ccatcaccgc cgcccaggct
     1741 tgttacgacc ggatcatcta ctgcgacaca gacagcatcc acctgacagg caccgaggtg
     1801 cctgagatca tcaaggacat cgtggaccct aagaagctgg gctactgggc tcatgagtcc
     1861 accttcaaaa gagccaagta cctgagacag aagacctaca tccaggacat ctacatgaag
     1921 gaagttgacg gaaagctggt ggagtgcagc cctgatgagg ccaccacaac caagttctcc
     1981 gtgaagtgcg ccggcatgac agataccatc aaaaagaagg tgacattcga caacttcaag
     2041 gtgggcttca gcagcaaggg aaagcctaaa ccagtgcaag tgaacggcgg agtggtgctg
     2101 gtagacagcg tgttcacaat caagtccggc ggaagctctg gtggcagcaa gcggaccgcc
     2161 gacggctctg aattcgagag ccctaagaag aaaagaaagg tgtgaccggt catcatcacc
     2221 atcaccattg agtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     2281 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     2341 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     2401 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     2461 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tcgataccgt
     2521 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     2581 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctagggtg
     2641 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2701 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2761 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2821 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2881 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2941 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3001 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3061 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3121 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3181 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3241 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3301 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3361 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3421 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3481 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3541 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3601 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3661 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3721 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3781 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3841 caatgatacc gcgcgaacca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3901 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3961 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4021 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4081 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4141 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4201 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4261 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4321 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4381 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4441 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4501 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4561 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4621 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4681 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcgatct cccgatcccc
     4741 tagggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     4801 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     4861 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
     4921 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat
     4981 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     5041 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     5101 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     5161 ggtaaactgc ccacttggca gtacatcaag tgtatc
//
