LOCUS       sequence-349376-        4498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human RNF216 at the 
            C-terminus with halotag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4498)
  TITLE     iNDI donor vectors for endogenous tagging with Halotag
REFERENCE   2  (bases 1 to 4498)
  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: RNF216_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    105..126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(141..146,147..164,165..171)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    179..195
                     /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)."
     primer_bind     184..206
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     203..219
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     203..219
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     280..296
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             748..1635
                     /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(2101..2117)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2110..2132)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2326..2345)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2445..2467
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2505..2523)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2591..2695
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2696..2764,2765..3556)
                     /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     complement(2914..2933)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3727..4315
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4216..4235
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4469..4486
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
       61 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
      121 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      181 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttgca
      241 tgcaggcctc tgcagtcgac gggcccggga tccgatgttg taaaacgacg gccagtgaat
      301 tcagaggtcg ctgccctggt cccaggctag attccattga ggttggttta atgtttttct
      361 ccactaatcc ccacatccct ttcagagaac accttcaaac gcattggacc cccgctggag
      421 aagcctgtgg agaaggtgca gagggtggag gccctcccga ggcccgttcc gcagaacctg
      481 ccacagccac agatgccacc ctatgccttc gcgcacccac ccttccccct gcctcccgtg
      541 cggcctgtgt tcaacaactt cccactcaac atggggccta tcccagcccc gtacgtgccc
      601 cctctgccca acgtgcgggt caactatgac ttcggtccca tccacatgcc cctggagcac
      661 aacctgccca tgcactttgg cccccagccg cggcatcgct tcggtggagg aggttctgga
      721 ggcggtggaa gtggtggcgg aggtagcgcc gagatcggca ccggctttcc cttcgatcct
      781 cactacgtgg aagtgctggg cgagagaatg cactatgtgg acgtgggccc cagagatgga
      841 acccctgtgc tgtttctgca cggcaaccct accagcagct acgtgtggcg gaacatcatc
      901 cctcacgtgg cccctacaca cagatgtatc gcccctgacc tgatcggcat gggcaagagc
      961 gataagcccg acctgggcta cttcttcgac gaccacgtgc ggttcatgga cgcctttatt
     1021 gaggccctcg gcctggaaga ggtggtgctg gttattcacg attggggcag cgccctggga
     1081 ttccactggg ccaagagaaa ccccgagaga gtgaagggaa tcgccttcat ggaattcatc
     1141 aggcccattc ctacctggga cgagtggcct gagttcgcca gagagacatt ccaggccttc
     1201 agaaccaccg acgtgggcag aaagctgatc atcgaccaga acgtgttcat cgagggcacc
     1261 ctgcctatgg gagtcgtcag accactgacc gaggtggaaa tggaccacta cagagagccc
     1321 tttctgaacc ccgtggaccg ggaacctctt tggagattcc ccaacgagct gcccattgcc
     1381 ggcgagcctg ctaatattgt ggccctggtg gaagagtaca tggactggct gcatcagagc
     1441 cccgtgccta agctgctgtt ttggggaaca cccggcgtgc tgattcctcc tgctgaagct
     1501 gccagactgg ccaagagcct gcctaattgc aaggccgtgg atatcggccc tggcctgaat
     1561 ctgctgcaag aggacaaccc cgatctgatc ggctccgaga ttgccagatg gctgagcacc
     1621 ctggaaatca gcggataatg gccccgaatc cccattgagc agcacaaagc ccgtttgggg
     1681 taggagtgtg gatggagaac cctcccccaa ggctggtgtc tgtaccattg catcctaagt
     1741 cagcttgaag ggtaggctgg ttttcttccc acccctttcc tagaagggct actgctcctg
     1801 gaagagtgga cggatccata ataaagacgt cccaaatggt ggagttcgga gagagctgcg
     1861 atgtgaactg cccctcccct cgcatccccc aggccaccaa cggcagtcct tctgccttgt
     1921 ccatggcata ggccatagac caggtccctg ctgctcacac ctgggcctct cctcggagcc
     1981 gacccctggg tagcaaggca gccgagagca tctccctgga ggggcccacg gttgggccaa
     2041 gcttggcgta atcatggtca tagcatctag atgcattcgc gaggtaccga gctcgaattc
     2101 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     2161 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     2221 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
     2281 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
     2341 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     2401 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     2461 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     2521 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     2581 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2641 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     2701 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2761 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2821 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2881 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     2941 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3001 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3061 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     3121 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3181 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     3241 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3301 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3361 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg gatctcgcgg
     3421 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3481 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3541 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3601 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3661 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3721 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3781 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3841 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3901 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3961 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4021 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4081 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4141 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4201 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4261 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4321 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4381 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4441 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagag
//
