LOCUS       sequence-349354-        4377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human OTUD4 at the C-terminus
            with halotag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4377)
  TITLE     iNDI donor vectors for endogenous tagging with Halotag
REFERENCE   2  (bases 1 to 4377)
  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: OTUD4_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             344..1231
                     /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(1653..1669)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1653..1669)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1666..1688)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1677..1693
                     /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(1701..1707,1708..1725,1726..1731))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1746..1767
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1884..1901)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2055..2643)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2135..2154)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2814..3605,3606..3674))
                     /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     3437..3456
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3675..3779)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3847..3865
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3903..3925)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4025..4044
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4238..4260
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4252..4269
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4253..4269
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 agtgcaaaga ccaaaagaag agagttcaga agatgaaaat gaagtgtcta atattttgag
       61 aagtggtaga tccaagcagt tctataatca aacttatgga agcaggaagt acaaaagtga
      121 ttggggctat tctggtaggg gtggatatca acatgtgaga agtgaggagt cctggaaagg
      181 acagccaagc agaagtcggg atgaaggtta tcagtaccat cgaaatgtca gagggcgacc
      241 atttagggga gataggagga gatcagggat gggagatggc cataggggac agcacactgg
      301 tggaggaggt tctggaggcg gtggaagtgg tggcggaggt agcgccgaga tcggcaccgg
      361 ctttcccttc gatcctcact acgtggaagt gctgggcgag agaatgcact atgtggacgt
      421 gggccccaga gatggaaccc ctgtgctgtt tctgcacggc aaccctacca gcagctacgt
      481 gtggcggaac atcatccctc acgtggcccc tacacacaga tgtatcgccc ctgacctgat
      541 cggcatgggc aagagcgata agcccgacct gggctacttc ttcgacgacc acgtgcggtt
      601 catggacgcc tttattgagg ccctcggcct ggaagaggtg gtgctggtta ttcacgattg
      661 gggcagcgcc ctgggattcc actgggccaa gagaaacccc gagagagtga agggaatcgc
      721 cttcatggaa ttcatcaggc ccattcctac ctgggacgag tggcctgagt tcgccagaga
      781 gacattccag gccttcagaa ccaccgacgt gggcagaaag ctgatcatcg accagaacgt
      841 gttcatcgag ggcaccctgc ctatgggagt cgtcagacca ctgaccgagg tggaaatgga
      901 ccactacaga gagccctttc tgaaccccgt ggaccgggaa cctctttgga gattccccaa
      961 cgagctgccc attgccggcg agcctgctaa tattgtggcc ctggtggaag agtacatgga
     1021 ctggctgcat cagagccccg tgcctaagct gctgttttgg ggaacacccg gcgtgctgat
     1081 tcctcctgct gaagctgcca gactggccaa gagcctgcct aattgcaagg ccgtggatat
     1141 cggccctggc ctgaatctgc tgcaagagga caaccccgat ctgatcggct ccgagattgc
     1201 cagatggctg agcaccctgg aaatcagcgg ataatggttg ttgccgaagt attttctaac
     1261 agaaactctt aggtggaatg tttctgaagg ctttaaaaaa aatacaataa agtaaaaacc
     1321 gcagttggaa ctctttcctc tccccagccc cccttacctt cactcccata ctggacaaga
     1381 cattttggac atgagttcaa gggggcaggt taattcctgg cattgccagt tttcttcatt
     1441 cagaatctgt ttgttgatca agtgactgtc cccgtaaaaa gtagaagata agtttgttaa
     1501 agtatttttt ataaacagct taatataaaa cattatagat ttagtgtttg gcttttttcc
     1561 ttataagttt ataagtttga tttcagatgt tggaaatgtg tgctttatag tgtttactaa
     1621 agtgacaaga aaataagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     1681 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     1741 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     1801 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     1861 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     1921 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     1981 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2041 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2101 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2161 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2221 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2281 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2341 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2401 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2461 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     2521 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2581 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2641 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2701 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2761 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2821 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     2881 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     2941 gcaatgatac cgcgagatcc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3001 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3061 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3121 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3181 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3241 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3301 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     3361 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     3421 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     3481 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     3541 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     3601 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3661 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3721 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3781 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     3841 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     3901 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     3961 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     4021 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     4081 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     4141 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     4201 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     4261 acggccagtg aattctaaaa ggaccattca aagcctgaaa gaaaaaacag aaaaagtaaa
     4321 agatcctaag actgctgctg atgtggtcag ccctggggcc aactctgttg atagcag
//
