LOCUS       sequence-349377-        4498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human PARK7 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: PARK7_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    95..116
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(131..136,137..154,155..161)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    169..185
                     /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     174..196
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     193..209
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     193..209
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     270..286
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             738..1625
                     /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(2091..2107)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2100..2122)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2316..2335)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2435..2457
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2495..2513)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2581..2685
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2686..2754,2755..3546)
                     /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(2904..2923)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3717..4305
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4206..4225
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4459..4476
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
       61 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      121 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
      181 aacaatttca cacaggaaac agctatgacc atgattacgc caagcttgca tgcaggcctc
      241 tgcagtcgac gggcccggga tccgatgttg taaaacgacg gccagtgaat tcgcatttaa
      301 ctcacgcata tttgtttcat tctaacagtg gtttctgtca ccctttgctc tgcacagttt
      361 taaaaatacc tttgtagggg gcttctaaga gcttggagtg cctagtaaat gtttttgaat
      421 ggttagctac agtgttgggt ttatatgctg taatagtgaa tttaattggt aagtaatcgt
      481 ctttctcgtc acatagccca ttaggatgtc accttttctg tttctacttt gcaggtcatt
      541 acacctactc tgagaatcgt gtggaaaaag acggcctgat tcttacaagc cgggggcctg
      601 ggaccagctt cgagtttgcg cttgcaattg ttgaagccct gaatggcaag gaggtggcgg
      661 ctcaagtgaa ggctccactt gttcttaaag acggtggagg aggttctgga ggcggtggaa
      721 gtggtggcgg aggtagcgcc gagatcggca ccggctttcc cttcgatcct cactacgtgg
      781 aagtgctggg cgagagaatg cactatgtgg acgtgggccc cagagatgga acccctgtgc
      841 tgtttctgca cggcaaccct accagcagct acgtgtggcg gaacatcatc cctcacgtgg
      901 cccctacaca cagatgtatc gcccctgacc tgatcggcat gggcaagagc gataagcccg
      961 acctgggcta cttcttcgac gaccacgtgc ggttcatgga cgcctttatt gaggccctcg
     1021 gcctggaaga ggtggtgctg gttattcacg attggggcag cgccctggga ttccactggg
     1081 ccaagagaaa ccccgagaga gtgaagggaa tcgccttcat ggaattcatc aggcccattc
     1141 ctacctggga cgagtggcct gagttcgcca gagagacatt ccaggccttc agaaccaccg
     1201 acgtgggcag aaagctgatc atcgaccaga acgtgttcat cgagggcacc ctgcctatgg
     1261 gagtcgtcag accactgacc gaggtggaaa tggaccacta cagagagccc tttctgaacc
     1321 ccgtggaccg ggaacctctt tggagattcc ccaacgagct gcccattgcc ggcgagcctg
     1381 ctaatattgt ggccctggtg gaagagtaca tggactggct gcatcagagc cccgtgccta
     1441 agctgctgtt ttggggaaca cccggcgtgc tgattcctcc tgctgaagct gccagactgg
     1501 ccaagagcct gcctaattgc aaggccgtgg atatcggccc tggcctgaat ctgctgcaag
     1561 aggacaaccc cgatctgatc ggctccgaga ttgccagatg gctgagcacc ctggaaatca
     1621 gcggataaag cagcgaactg cgacgatcac ttagagaaac aggccgttag gaatccattc
     1681 tcactgtgtt cgctctaaac aaaacagtgg taggttaatg tgttcagaag tcgctgtcct
     1741 tactactttt gcggaagtat ggaagtcaca actacacaga gatttctcag cctacaaatt
     1801 gtgtctatac atttctaagc cttgtttgca gaataaacag ggcatttagc aaactactga
     1861 ttgtttcttg ttttgtctct catttctttt gtgaaattaa attccgtatc accttcattt
     1921 gcagctctta actgtccata tggcactgaa ataaaagaac agtgaccaca ttttacacag
     1981 caaggaggaa aggcatacaa acagaattta agaggcttgt gattttctaa gcttggcgta
     2041 atcatggtca tagcatctag atgcattcgc gaggtaccga gctcgaattc actggccgtc
     2101 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     2161 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     2221 cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
     2281 tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag
     2341 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     2401 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     2461 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     2521 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     2581 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2641 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     2701 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     2761 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     2821 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     2881 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     2941 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3001 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     3061 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     3121 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     3181 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     3241 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     3301 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     3361 ggctggttta ttgctgataa atctggagcc ggtgagcgtg gatctcgcgg tatcattgca
     3421 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     3481 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     3541 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     3601 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     3661 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3721 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3781 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3841 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     3901 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3961 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4021 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4081 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4141 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4201 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4261 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4321 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     4381 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     4441 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatac
//
