LOCUS       sequence-349381-        4377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human PLA2G6 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: PLA2G6_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(528..1319,1320..1388))
                     /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     1151..1170
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1389..1493)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1561..1579
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1617..1639)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1739..1758
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1952..1974
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1966..1983
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1967..1983
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2435..3322
                     /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(3744..3760)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3744..3760)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3757..3779)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3768..3784
                     /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(3792..3798,3799..3816,3817..3822))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3837..3858
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3975..3992)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(4146..4377,1..357))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4226..4245)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
       61 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
      121 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
      181 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
      241 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
      301 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
      361 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
      421 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
      481 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
      541 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
      601 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
      661 ataccgcgag atccacgctc accggctcca gatttatcag caataaacca gccagccgga
      721 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
      781 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
      841 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
      901 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
      961 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     1021 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     1081 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     1141 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     1201 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     1261 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     1321 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     1381 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     1441 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     1501 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     1561 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     1621 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     1681 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     1741 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     1801 gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa
     1861 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     1921 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     1981 agtgaattcc gggcgggctg tggaccgggc acgggcctgg tgcgagatgg tcggcatcca
     2041 gtacttcagg tgagggctca gccgccccag cccttggccc cgtgccctgg cgtggtcgga
     2101 ctcaccgacc ttcccctccc agccctagtg tggactttcc cttccccaag ggtctgctct
     2161 gttccccaag cccaccctgg tcctagctgg cgtcccctgc ccagcctgag catcctaggg
     2221 tgaccccctc ctccctggcc cgaacagatt gaacccccag ctggggacgg acatcatgct
     2281 ggatgaggtc agtgacacag tgctggtcaa cgccctctgg gagaccgagg tctacatcta
     2341 tgagcaccgc gaggagttcc agaagctcat ccagctgctg ctctcacccg gtggaggagg
     2401 ttctggaggc ggtggaagtg gtggcggagg tagcgccgag atcggcaccg gctttccctt
     2461 cgatcctcac tacgtggaag tgctgggcga gagaatgcac tatgtggacg tgggccccag
     2521 agatggaacc cctgtgctgt ttctgcacgg caaccctacc agcagctacg tgtggcggaa
     2581 catcatccct cacgtggccc ctacacacag atgtatcgcc cctgacctga tcggcatggg
     2641 caagagcgat aagcccgacc tgggctactt cttcgacgac cacgtgcggt tcatggacgc
     2701 ctttattgag gccctcggcc tggaagaggt ggtgctggtt attcacgatt ggggcagcgc
     2761 cctgggattc cactgggcca agagaaaccc cgagagagtg aagggaatcg ccttcatgga
     2821 attcatcagg cccattccta cctgggacga gtggcctgag ttcgccagag agacattcca
     2881 ggccttcaga accaccgacg tgggcagaaa gctgatcatc gaccagaacg tgttcatcga
     2941 gggcaccctg cctatgggag tcgtcagacc actgaccgag gtggaaatgg accactacag
     3001 agagcccttt ctgaaccccg tggaccggga acctctttgg agattcccca acgagctgcc
     3061 cattgccggc gagcctgcta atattgtggc cctggtggaa gagtacatgg actggctgca
     3121 tcagagcccc gtgcctaagc tgctgttttg gggaacaccc ggcgtgctga ttcctcctgc
     3181 tgaagctgcc agactggcca agagcctgcc taattgcaag gccgtggata tcggccctgg
     3241 cctgaatctg ctgcaagagg acaaccccga tctgatcggc tccgagattg ccagatggct
     3301 gagcaccctg gaaatcagcg gataagggtc cccagcctct caccggcccc agctgacctc
     3361 gtccattcag cccctgccag gccaagccca gccactgccc tcccgggcag atctgggccc
     3421 aggcacctct gagtccatag accaggcctg ggagaatgcc aagctgcctg cccgaggctg
     3481 gtcctgaagg cctgtctccc actaaccccg ccttccagca ctttctgtca ttccaggctg
     3541 ggaaagtcta gagccccctt tggccccttt ccctgactgt caaggacaac tgactccccc
     3601 atcagctcaa acattaaggg tacccgggca caaccgtacc cctgccccca gccccagcct
     3661 ccctgagggc ctgccgggct gcctctgccc cagcccccag caagggcact cccaggcttc
     3721 ctggtaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     3781 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     3841 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     3901 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3961 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4021 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4081 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4141 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4201 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4261 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4321 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtc
//
