LOCUS       sequence-349363-        4377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human VAPB 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: VAPB_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(162..750)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(242..261)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(921..1712,1713..1781))
                     /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     1544..1563
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1782..1886)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1954..1972
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2010..2032)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2132..2151
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2345..2367
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2359..2376
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2360..2376
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2828..3715
                     /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(4137..4153)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4137..4153)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4150..4172)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4161..4177
                     /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(4185..4191,4192..4209,4210..4215))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4230..4251
                     /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(4368..4377,1..8))
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
       61 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
      121 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
      181 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      241 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      301 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      361 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      421 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      481 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      541 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      601 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      661 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      721 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      781 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
      841 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
      901 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
      961 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     1021 gagggcttac catctggccc cagtgctgca atgataccgc gagatccacg ctcaccggct
     1081 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1141 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1201 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1261 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1321 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1381 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1441 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1501 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1561 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1621 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     1681 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     1741 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     1801 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     1861 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     1921 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     1981 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     2041 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     2101 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     2161 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     2221 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     2281 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     2341 ttttcccagt cacgacgttg taaaacgacg gccagtgaat tcgatgtctg ttgtaactag
     2401 cagcagcaag tggaatggat gaaagaaaag aggggactgt tttctcagtg gcttcaggct
     2461 agccctttcc cctctgtagc ctgcagtttc tccgtttgca aaatgcggtt ggaccatatc
     2521 atgtcatcca acactgggca taaacagccc atcccgttaa gctgtacagt tgactcccct
     2581 ttctggtgcc ttggcttgtc tttgaaatgt gcgtttgctc ccgtaggaag aagatggact
     2641 gcggatgagg aagacagtgc agagcaacag ccccatttca gcattagccc caactgggaa
     2701 ggaagaaggc cttagcaccc ggctcttggc tctggtggtt ttgttcttta tcgttggtgt
     2761 aattattggg aagattgcct tgggtggagg aggttctgga ggcggtggaa gtggtggcgg
     2821 aggtagcgcc gagatcggca ccggctttcc cttcgatcct cactacgtgg aagtgctggg
     2881 cgagagaatg cactatgtgg acgtgggccc cagagatgga acccctgtgc tgtttctgca
     2941 cggcaaccct accagcagct acgtgtggcg gaacatcatc cctcacgtgg cccctacaca
     3001 cagatgtatc gcccctgacc tgatcggcat gggcaagagc gataagcccg acctgggcta
     3061 cttcttcgac gaccacgtgc ggttcatgga cgcctttatt gaggccctcg gcctggaaga
     3121 ggtggtgctg gttattcacg attggggcag cgccctggga ttccactggg ccaagagaaa
     3181 ccccgagaga gtgaagggaa tcgccttcat ggaattcatc aggcccattc ctacctggga
     3241 cgagtggcct gagttcgcca gagagacatt ccaggccttc agaaccaccg acgtgggcag
     3301 aaagctgatc atcgaccaga acgtgttcat cgagggcacc ctgcctatgg gagtcgtcag
     3361 accactgacc gaggtggaaa tggaccacta cagagagccc tttctgaacc ccgtggaccg
     3421 ggaacctctt tggagattcc ccaacgagct gcccattgcc ggcgagcctg ctaatattgt
     3481 ggccctggtg gaagagtaca tggactggct gcatcagagc cccgtgccta agctgctgtt
     3541 ttggggaaca cccggcgtgc tgattcctcc tgctgaagct gccagactgg ccaagagcct
     3601 gcctaattgc aaggccgtgg atatcggccc tggcctgaat ctgctgcaag aggacaaccc
     3661 cgatctgatc ggctccgaga ttgccagatg gctgagcacc ctggaaatca gcggataaag
     3721 gtagcatgca caggatggta aattggattg gtggatccac catatcatgg gatttaaatt
     3781 tatcataacc atgtgtaaaa agaaattaat gtatgatgac atctcacagg tcttgccttt
     3841 aaattacccc tccctgcaca cacatacaca gatacacaca cacaaatata atgtaacgat
     3901 cttttagaaa gttaaaaatg tatagtaact gattgagggg gaaaagaatg atctttatta
     3961 atgacaaggg aaaccatgag taatgccaca atggcatatt gtaaatgtca ttttaaacat
     4021 tggtaggcct tggtacatga tgctggatta cctctcttaa aatgacaccc ttcctcgcct
     4081 gttggtgctg gcccttgggg agctggagcc cagcatgcaa gcttggcgta atcatggtca
     4141 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4201 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4261 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4321 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcact
//
