LOCUS       sequence-349328-        4377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human PLEKHG4 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: PLEKHG4_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(166..754)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(246..265)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(925..1716,1717..1785))
                     /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     1548..1567
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1786..1890)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1958..1976
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2014..2036)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2136..2155
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2349..2371
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2363..2380
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2364..2380
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2832..3719
                     /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(4141..4157)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4141..4157)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4154..4176)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4165..4181
                     /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(4189..4195,4196..4213,4214..4219))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4234..4255
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(join(4372..4377,1..12))
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
       61 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
      121 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
      181 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
      241 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
      301 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
      361 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
      421 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
      481 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
      541 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
      601 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
      661 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
      721 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
      781 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
      841 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
      901 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
      961 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     1021 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagatc cacgctcacc
     1081 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     1141 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     1201 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     1261 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     1321 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     1381 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     1441 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     1501 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     1561 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     1621 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     1681 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     1741 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     1801 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     1861 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     1921 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     1981 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     2041 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     2101 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     2161 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     2221 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     2281 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     2341 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattctctg ggcctagcca
     2401 gaggcaggag ggcatggctt ggggtctggg gcttcccgtg ctgacagttc acccagtctc
     2461 cctccctagc tgctgaggac tcagagatct cgtcccaatg cccatcagcc agtggctcca
     2521 gtggctctga cagcagctgt gtgtcagggc aggccctggg taggggcctg gaggacttac
     2581 cctgtgtgag tgccatttgc cctataccca ccagcccttc cccagcccaa gctgagcctg
     2641 tgactgtgac cagcccctcc ccagcccagg ctaggccttt actttgggta gagcttgggg
     2701 tgggtgggcc agagccattc cccagacacc tcctgggtcc agagtcaggg aagcaggcat
     2761 tcatgcctgg cctggccctg caggtcggtg gaggaggttc tggaggcggt ggaagtggtg
     2821 gcggaggtag cgccgagatc ggcaccggct ttcccttcga tcctcactac gtggaagtgc
     2881 tgggcgagag aatgcactat gtggacgtgg gccccagaga tggaacccct gtgctgtttc
     2941 tgcacggcaa ccctaccagc agctacgtgt ggcggaacat catccctcac gtggccccta
     3001 cacacagatg tatcgcccct gacctgatcg gcatgggcaa gagcgataag cccgacctgg
     3061 gctacttctt cgacgaccac gtgcggttca tggacgcctt tattgaggcc ctcggcctgg
     3121 aagaggtggt gctggttatt cacgattggg gcagcgccct gggattccac tgggccaaga
     3181 gaaaccccga gagagtgaag ggaatcgcct tcatggaatt catcaggccc attcctacct
     3241 gggacgagtg gcctgagttc gccagagaga cattccaggc cttcagaacc accgacgtgg
     3301 gcagaaagct gatcatcgac cagaacgtgt tcatcgaggg caccctgcct atgggagtcg
     3361 tcagaccact gaccgaggtg gaaatggacc actacagaga gccctttctg aaccccgtgg
     3421 accgggaacc tctttggaga ttccccaacg agctgcccat tgccggcgag cctgctaata
     3481 ttgtggccct ggtggaagag tacatggact ggctgcatca gagccccgtg cctaagctgc
     3541 tgttttgggg aacacccggc gtgctgattc ctcctgctga agctgccaga ctggccaaga
     3601 gcctgcctaa ttgcaaggcc gtggatatcg gccctggcct gaatctgctg caagaggaca
     3661 accccgatct gatcggctcc gagattgcca gatggctgag caccctggaa atcagcggat
     3721 aagcccggga ctggacgagc agtagatcca gcagcctgca gctccaagga acattgcctc
     3781 tctggatctg ctgtgaccag ggtgtggctg acacctgggc tacctccaac ctacatgtgc
     3841 aacgctgttg actacccttt ctgatgtgtg tggccattgg actaactggc acggggcctc
     3901 tctagggaag tctggttgta gagcctgaat aggctcctgg ccccatgacc ccttctcctg
     3961 tccccagctc ccatcccagt tgtgggttaa gaataggcta gagcagacat tgggtgtttc
     4021 catgctgtag gctggtgggg gaccatgtgc ctctaggcag tgactagggt gcccccaccc
     4081 ctcaggaaga acacaggtgg gctcctagca gctgatcccc aaaagcttgg cgtaatcatg
     4141 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4201 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4261 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4321 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgct
//
