LOCUS       sequence-349382-        4456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human DNMT1 at the C-terminus
            with halotag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4456)
  TITLE     iNDI donor vectors for endogenous tagging with Halotag
REFERENCE   2  (bases 1 to 4456)
  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"
COMMENT     Sequence Label: DNMT1_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(64..81)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(235..823)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(315..334)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(994..1785,1786..1854))
                     /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     1617..1636
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1855..1959)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2027..2045
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2083..2105)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2205..2224
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2418..2440
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2432..2449
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2433..2449
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2936..3823
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="AEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     primer_bind     complement(4289..4305)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4289..4305)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4302..4324)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4313..4329
                     /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(4337..4343,4344..4361,4362..4367))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4382..4403
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
       61 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
      121 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
      181 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
      241 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
      301 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
      361 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
      421 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
      481 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
      541 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
      601 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
      661 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
      721 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
      781 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
      841 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
      901 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
      961 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     1021 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     1081 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagatcc
     1141 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     1201 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     1261 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     1321 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     1381 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     1441 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     1501 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     1561 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     1621 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     1681 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     1741 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     1801 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     1861 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     1921 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     1981 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     2041 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     2101 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     2161 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     2221 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     2281 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     2341 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     2401 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattcgagct
     2461 cggtacctcg cgaatgcatc tagatccaat tgtgaggatt gagtgagttg cacgtgtcaa
     2521 gtgcttagag caggcgtgct gcacacagca ggcctttggt caggttggct gctgggctgg
     2581 ccctggggcc gtttccctca ctcctgctcg gtgaatttgg ctcagcaggc acctgcctca
     2641 gctgctcact tgagcctctg ggtctagaac cctctgggga ccgtttgagg agtgttcagt
     2701 ctccgtgaac gttcccttag cactctgcca cttattgggt cagctgttaa catcagtacg
     2761 ttaatgtttc ctgatggtcc atgtctgtta ctcgcctgtc aagtggcgtg acaccgggcg
     2821 tgttccccag agtgactttt ccttttattt cccttcagct aaaataaagg aggaggaagc
     2881 tgctaaggac ggtggaggag gttctggagg cggtggaagt ggtggcggag gtagcgccga
     2941 gatcggcacc ggctttccct tcgatcctca ctacgtggaa gtgctgggcg agagaatgca
     3001 ctatgtggac gtgggcccca gagatggaac ccctgtgctg tttctgcacg gcaaccctac
     3061 cagcagctac gtgtggcgga acatcatccc tcacgtggcc cctacacaca gatgtatcgc
     3121 ccctgacctg atcggcatgg gcaagagcga taagcccgac ctgggctact tcttcgacga
     3181 ccacgtgcgg ttcatggacg cctttattga ggccctcggc ctggaagagg tggtgctggt
     3241 tattcacgat tggggcagcg ccctgggatt ccactgggcc aagagaaacc ccgagagagt
     3301 gaagggaatc gccttcatgg aattcatcag gcccattcct acctgggacg agtggcctga
     3361 gttcgccaga gagacattcc aggccttcag aaccaccgac gtgggcagaa agctgatcat
     3421 cgaccagaac gtgttcatcg agggcaccct gcctatggga gtcgtcagac cactgaccga
     3481 ggtggaaatg gaccactaca gagagccctt tctgaacccc gtggaccggg aacctctttg
     3541 gagattcccc aacgagctgc ccattgccgg cgagcctgct aatattgtgg ccctggtgga
     3601 agagtacatg gactggctgc atcagagccc cgtgcctaag ctgctgtttt ggggaacacc
     3661 cggcgtgctg attcctcctg ctgaagctgc cagactggcc aagagcctgc ctaattgcaa
     3721 ggccgtggat atcggccctg gcctgaatct gctgcaagag gacaaccccg atctgatcgg
     3781 ctccgagatt gccagatggc tgagcaccct ggaaatcagc ggataattct gccctcccgt
     3841 cacccctgtt tctcgcacca ggaatcccca acatgcactg atgttgtgtt tttaacatgt
     3901 caatctgtcc gttcacatgt gtggtacatg gtgtttgtgg ccttggctga catgaagctg
     3961 ttgtgtgagg ttcgcttatc aactaatgat ttagtgatca aattgtgcag tactttgtgc
     4021 attctggatt ttaaaagttt tttattatgc attatatcaa atctaccact gtatgagtgg
     4081 aaattaagac tttatgtagt ttttatatgt tgtaatattt cttcaaataa atctctccta
     4141 taaaccaccc tgggtgggta ttcgtgattt gcacgggact tctggctgag gtcagacagg
     4201 aacaagctct tccatgctga aggcgaccaa tatcggatcc cgggcccgtc gactgcagag
     4261 gcctgcatgc aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4321 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4381 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4441 acctgtcgtg ccagct
//
