LOCUS       sequence-349343-        4377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human POLG 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: POLG_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(21..38)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(192..780)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(272..291)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(951..1742,1743..1811))
                     /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     1574..1593
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1812..1916)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1984..2002
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2040..2062)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2162..2181
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2375..2397
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2389..2406
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2390..2406
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2858..3745
                     /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(4167..4183)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4167..4183)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4180..4202)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4191..4207
                     /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(4215..4221,4222..4239,4240..4245))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4260..4281
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
       61 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
      121 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
      181 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
      241 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
      301 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
      361 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
      421 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
      481 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
      541 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
      601 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
      661 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
      721 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
      781 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
      841 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      901 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      961 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     1021 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     1081 atgataccgc gagatccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     1141 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     1201 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     1261 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     1321 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     1381 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     1441 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     1501 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     1561 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     1621 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     1681 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     1741 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1801 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1861 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1921 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     1981 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     2041 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     2101 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     2161 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     2221 tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg
     2281 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     2341 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     2401 gccagtgaat tccccctact tcaaccacag aagaaatgtt taaagataat ttattctata
     2461 gagtgcattc ttgcactcta taggtgacag aaaaacaaac tgtgctttaa ataccaaaca
     2521 agtaaatcag aaagcttatt ttctatttaa aatatatcta agacacactt atataaaaag
     2581 aaaacagacc ctcctaacat gtaacattac cgttcgtggc aattgttctc aacctttcac
     2641 tctccttttg accttagcat taagctcctt tgctcacttc tgagctctca gttacagttc
     2701 ttgaggtggc atcctaacca atttgcacta tctttcaggt gaagcgctgg atatttacca
     2761 gataattgaa ctcaccaaag gctccttgga aaaacgaagc cagcctggac caggtggagg
     2821 aggttctgga ggcggtggaa gtggtggcgg aggtagcgcc gagatcggca ccggctttcc
     2881 cttcgatcct cactacgtgg aagtgctggg cgagagaatg cactatgtgg acgtgggccc
     2941 cagagatgga acccctgtgc tgtttctgca cggcaaccct accagcagct acgtgtggcg
     3001 gaacatcatc cctcacgtgg cccctacaca cagatgtatc gcccctgacc tgatcggcat
     3061 gggcaagagc gataagcccg acctgggcta cttcttcgac gaccacgtgc ggttcatgga
     3121 cgcctttatt gaggccctcg gcctggaaga ggtggtgctg gttattcacg attggggcag
     3181 cgccctggga ttccactggg ccaagagaaa ccccgagaga gtgaagggaa tcgccttcat
     3241 ggaattcatc aggcccattc ctacctggga cgagtggcct gagttcgcca gagagacatt
     3301 ccaggccttc agaaccaccg acgtgggcag aaagctgatc atcgaccaga acgtgttcat
     3361 cgagggcacc ctgcctatgg gagtcgtcag accactgacc gaggtggaaa tggaccacta
     3421 cagagagccc tttctgaacc ccgtggaccg ggaacctctt tggagattcc ccaacgagct
     3481 gcccattgcc ggcgagcctg ctaatattgt ggccctggtg gaagagtaca tggactggct
     3541 gcatcagagc cccgtgccta agctgctgtt ttggggaaca cccggcgtgc tgattcctcc
     3601 tgctgaagct gccagactgg ccaagagcct gcctaattgc aaggccgtgg atatcggccc
     3661 tggcctgaat ctgctgcaag aggacaaccc cgatctgatc ggctccgaga ttgccagatg
     3721 gctgagcacc ctggaaatca gcggataaca ctgcctggag gctctgtatt tgctcccgtg
     3781 gagcttcatc ggggtggtgc aggctcccaa actcaggctt tcagctgtgc tttttgcaaa
     3841 agggcttgcc taaggccagc catttttcag tagcaggacc tgccaagaag attccttcta
     3901 actgaaggtg cagttgaatt cagtgggttc agaaccaaga tgccaacatc ggtgtggact
     3961 acaggacaag gggcattgtt gcttgttggg taaaaatgaa gcagaagccc caaagttcac
     4021 attaactcag gcatttcatt tattttttcc ttttcttctt ggctggttct ttgttctgtc
     4081 ccccatgctc tgatgcagtg ccctagaagg ggaaagaatt aatgctctaa cgtgataaac
     4141 ctgctccaaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4201 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4261 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4321 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgt
//
