LOCUS       sequence-349374-        4442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human APP at the C-terminus 
            with halotag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4442)
  TITLE     iNDI donor vectors for endogenous tagging with Halotag
REFERENCE   2  (bases 1 to 4442)
  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: APP_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(32..49)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(203..791)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(283..302)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(962..1753,1754..1822))
                     /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     1585..1604
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1823..1927)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1995..2013
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2051..2073)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2173..2192
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2386..2408
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2400..2417
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2401..2417
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2898..3785
                     /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(4243..4259)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4243..4259)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4256..4278)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4267..4283
                     /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(4291..4297,4298..4315,4316..4321))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4336..4357
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
       61 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
      121 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
      181 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
      241 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
      301 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
      361 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
      421 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
      481 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
      541 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
      601 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
      661 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
      721 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
      781 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
      841 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
      901 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
      961 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     1021 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     1081 ccagtgctgc aatgataccg cgagatccac gctcaccggc tccagattta tcagcaataa
     1141 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     1201 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     1261 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     1321 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     1381 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     1441 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     1501 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     1561 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     1621 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     1681 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     1741 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     1801 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     1861 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     1921 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     1981 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     2041 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     2101 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     2161 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     2221 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
     2281 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     2341 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     2401 gtaaaacgac ggccagtgaa ttcgagctcg gtacctcgcg aatgcatcta gaaccattgc
     2461 actgggttta tccacttact gtggagttgg ggaaccagaa acaaccactg gacactttgc
     2521 aagcgcgtca aggtttccac cgccacctgg tggctagatg gagcacagca tctgtgtaac
     2581 tggtgggcaa aggcgttctg ctccaagatg tcaaagtggg ggagaaaatt atgggtgttc
     2641 tgcaatcttg gcaaaagaag agtactgtac tccttatctt tttactgctt ctccatgttc
     2701 acccttaaaa gattgatttt tattttttac tcagctctcc tcttgttttt caggttgacg
     2761 ccgctgtcac cccagaggag cgccacctgt ccaagatgca gcagaacggc tacgaaaatc
     2821 caacctacaa gttctttgag cagatgcaga acggtggagg aggttctgga ggcggtggaa
     2881 gtggtggcgg aggtagcgcc gagatcggca ccggctttcc cttcgatcct cactacgtgg
     2941 aagtgctggg cgagagaatg cactatgtgg acgtgggccc cagagatgga acccctgtgc
     3001 tgtttctgca cggcaaccct accagcagct acgtgtggcg gaacatcatc cctcacgtgg
     3061 cccctacaca cagatgtatc gcccctgacc tgatcggcat gggcaagagc gataagcccg
     3121 acctgggcta cttcttcgac gaccacgtgc ggttcatgga cgcctttatt gaggccctcg
     3181 gcctggaaga ggtggtgctg gttattcacg attggggcag cgccctggga ttccactggg
     3241 ccaagagaaa ccccgagaga gtgaagggaa tcgccttcat ggaattcatc aggcccattc
     3301 ctacctggga cgagtggcct gagttcgcca gagagacatt ccaggccttc agaaccaccg
     3361 acgtgggcag aaagctgatc atcgaccaga acgtgttcat cgagggcacc ctgcctatgg
     3421 gagtcgtcag accactgacc gaggtggaaa tggaccacta cagagagccc tttctgaacc
     3481 ccgtggaccg ggaacctctt tggagattcc ccaacgagct gcccattgcc ggcgagcctg
     3541 ctaatattgt ggccctggtg gaagagtaca tggactggct gcatcagagc cccgtgccta
     3601 agctgctgtt ttggggaaca cccggcgtgc tgattcctcc tgctgaagct gccagactgg
     3661 ccaagagcct gcctaattgc aaggccgtgg atatcggccc tggcctgaat ctgctgcaag
     3721 aggacaaccc cgatctgatc ggctccgaga ttgccagatg gctgagcacc ctggaaatca
     3781 gcggataaac ccccgccaca gcagcgtctg aagttggaca gcaaaaccat tgcttcacta
     3841 cccatcggtg tccatttata gaataatgtg ggaagaaaca aacccgtttt atgatttact
     3901 cattatcgcc ttttgacagc tgtgctgtaa cacaagtaga tgcctgaact tgaattaatc
     3961 cacacatcag taatgtattc tatctctctt tacattttgg tctctatact acattattaa
     4021 tgggttttgt gtactgtaaa gaatttagct gtatcaaact agtgcatgaa tagattctct
     4081 cctgattatt tatcacatag ccccttagcc agttgtatat tattcttgtg gtttgtgacc
     4141 caattaagtc ctactttaca tatgctttaa gaatcgatgg gggatgctgg atcccgggcc
     4201 cgtcgactgc agaggcctgc atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt
     4261 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4321 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4381 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4441 ag
//
