LOCUS       sequence-349364-        4377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human PRNP 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: PRNP_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(98..889,890..958))
                     /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     721..740
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(959..1063)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1131..1149
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1187..1209)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1309..1328
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1522..1544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1536..1553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1537..1553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2005..2892
                     /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(3314..3330)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3314..3330)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3327..3349)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3338..3354
                     /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(3362..3368,3369..3386,3387..3392))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3407..3428
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3545..3562)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3716..4304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3796..3815)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
       61 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
      121 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
      181 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
      241 atccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
      301 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
      361 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
      421 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
      481 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
      541 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
      601 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
      661 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
      721 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
      781 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
      841 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
      901 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
      961 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     1021 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     1081 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     1141 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     1201 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     1261 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     1321 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     1381 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     1441 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     1501 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattca
     1561 gtggtggggg gccttggcgg ctacatgctg ggaagtgcca tgagcaggcc catcatacat
     1621 ttcggcagtg actatgagga ccgttactat cgtgaaaaca tgcaccgtta ccccaaccaa
     1681 gtgtactaca ggcccatgga tgagtacagc aaccagaaca actttgtgca cgactgcgtc
     1741 aatatcacaa tcaagcagca cacggtcacc acaaccacca agggggagaa cttcaccgag
     1801 accgacgtta agatgatgga gcgcgtggtt gagcagatgt gtatcaccca gtacgagagg
     1861 gaatctcagg cctattacca gagaggatcg agcatggtcc tcttctcctc tccacctgtg
     1921 atcctcctga tctctttcct catcttcctg atagtgggag gtggaggagg ttctggaggc
     1981 ggtggaagtg gtggcggagg tagcgccgag atcggcaccg gctttccctt cgatcctcac
     2041 tacgtggaag tgctgggcga gagaatgcac tatgtggacg tgggccccag agatggaacc
     2101 cctgtgctgt ttctgcacgg caaccctacc agcagctacg tgtggcggaa catcatccct
     2161 cacgtggccc ctacacacag atgtatcgcc cctgacctga tcggcatggg caagagcgat
     2221 aagcccgacc tgggctactt cttcgacgac cacgtgcggt tcatggacgc ctttattgag
     2281 gccctcggcc tggaagaggt ggtgctggtt attcacgatt ggggcagcgc cctgggattc
     2341 cactgggcca agagaaaccc cgagagagtg aagggaatcg ccttcatgga attcatcagg
     2401 cccattccta cctgggacga gtggcctgag ttcgccagag agacattcca ggccttcaga
     2461 accaccgacg tgggcagaaa gctgatcatc gaccagaacg tgttcatcga gggcaccctg
     2521 cctatgggag tcgtcagacc actgaccgag gtggaaatgg accactacag agagcccttt
     2581 ctgaaccccg tggaccggga acctctttgg agattcccca acgagctgcc cattgccggc
     2641 gagcctgcta atattgtggc cctggtggaa gagtacatgg actggctgca tcagagcccc
     2701 gtgcctaagc tgctgttttg gggaacaccc ggcgtgctga ttcctcctgc tgaagctgcc
     2761 agactggcca agagcctgcc taattgcaag gccgtggata tcggccctgg cctgaatctg
     2821 ctgcaagagg acaaccccga tctgatcggc tccgagattg ccagatggct gagcaccctg
     2881 gaaatcagcg gataaggaag gtcttcctgt tttcaccatc tttctaatct ttttccagct
     2941 tgagggaggc ggtatccacc tgcagccctt ttagtggtgg tgtctcactc tttcttctct
     3001 ctttgtcccg gataggctaa tcaataccct tggcactgat gggcactgga aaacatagag
     3061 tagacctgag atgctggtca agcccccttt gattgagttc atcatgagcc gttgctaatg
     3121 ccaggccagt aaaagtataa cagcaaataa ccattggtta atctggactt atttttggac
     3181 ttagtgcaac aggttgaggc taaaacaaat ctcagaacag tctgaaatac ctttgcctgg
     3241 atacctctgg ctccttcagc agctagagct cagtatacta atgccctatc ttagtaagct
     3301 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3361 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3421 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3481 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3541 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3601 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3661 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3721 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3781 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3841 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3901 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3961 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4021 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4081 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4141 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4201 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4261 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4321 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtca
//
