LOCUS       sequence-349326-        4377 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Donor vector for endogenous tagging of human PSEN2 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 Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: PSEN2_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(35..52)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(206..794)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(286..305)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(965..1756,1757..1825))
                     /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     1588..1607
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1826..1930)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1998..2016
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2054..2076)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2176..2195
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2389..2411
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2403..2420
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2404..2420
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2872..3759
                     /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(4181..4197)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4181..4197)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4194..4216)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4205..4221
                     /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(4229..4235,4236..4253,4254..4259))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4274..4295
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
       61 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      121 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
      181 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      241 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      301 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      361 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
      421 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
      481 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
      541 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
      601 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
      661 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
      721 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
      781 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
      841 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
      901 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
      961 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1021 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1081 gccccagtgc tgcaatgata ccgcgagatc cacgctcacc ggctccagat ttatcagcaa
     1141 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1201 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1261 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1321 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     1381 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     1441 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     1501 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     1561 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     1621 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     1681 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     1741 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     1801 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     1861 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     1921 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     1981 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     2041 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     2101 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     2161 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     2221 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     2281 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     2341 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     2401 gttgtaaaac gacggccagt gaattcgctc caggagtcag gaggcctgcc ttctggttca
     2461 ctcactaaca ggtgaggtga tctaatgggg gtgagaactt ctgcccttaa cacctcaaga
     2521 gctgttgcag gaccagggaa gataatgggg tgtctagcgc cgttatccga ctggtcctcg
     2581 aacaagctcc tgtgcccagg gactagacca tgactcacag ctcctgtcca caccagggat
     2641 caccacgctc accctcccct ccatgtcctg cagggcttgt gtctgaccct cctgctgctt
     2701 gctgtgttca agaaggcgct gcccgccctc cccatctcca tcacgttcgg gctcatcttt
     2761 tacttctcca cggacaacct ggtgcggccg ttcatggaca ccctggcctc ccatcagctc
     2821 tacatcggtg gaggaggttc tggaggcggt ggaagtggtg gcggaggtag cgccgagatc
     2881 ggcaccggct ttcccttcga tcctcactac gtggaagtgc tgggcgagag aatgcactat
     2941 gtggacgtgg gccccagaga tggaacccct gtgctgtttc tgcacggcaa ccctaccagc
     3001 agctacgtgt ggcggaacat catccctcac gtggccccta cacacagatg tatcgcccct
     3061 gacctgatcg gcatgggcaa gagcgataag cccgacctgg gctacttctt cgacgaccac
     3121 gtgcggttca tggacgcctt tattgaggcc ctcggcctgg aagaggtggt gctggttatt
     3181 cacgattggg gcagcgccct gggattccac tgggccaaga gaaaccccga gagagtgaag
     3241 ggaatcgcct tcatggaatt catcaggccc attcctacct gggacgagtg gcctgagttc
     3301 gccagagaga cattccaggc cttcagaacc accgacgtgg gcagaaagct gatcatcgac
     3361 cagaacgtgt tcatcgaggg caccctgcct atgggagtcg tcagaccact gaccgaggtg
     3421 gaaatggacc actacagaga gccctttctg aaccccgtgg accgggaacc tctttggaga
     3481 ttccccaacg agctgcccat tgccggcgag cctgctaata ttgtggccct ggtggaagag
     3541 tacatggact ggctgcatca gagccccgtg cctaagctgc tgttttgggg aacacccggc
     3601 gtgctgattc ctcctgctga agctgccaga ctggccaaga gcctgcctaa ttgcaaggcc
     3661 gtggatatcg gccctggcct gaatctgctg caagaggaca accccgatct gatcggctcc
     3721 gagattgcca gatggctgag caccctggaa atcagcggat aagggacatg gtgtgccaca
     3781 ggctgcaagc tgcagggaat tttcattgga tgcagttgta tagttttaca ctctagtgcc
     3841 atatattttt aagacttttc tttccttaaa aaataaagta cgtgtttact tggtgaggag
     3901 gaggcagaac cagctctttg gtgccagctg tttcatcacc agactttggc tcccgctttg
     3961 gggagcgcct cgcttcacgg acaggaagca cagcaggttt atccagatga actgagaagg
     4021 tcagattagg ggggggagaa gagcatccgg catgagggct gagatgcgca aagagtgtgc
     4081 tcgggagtgg cccctggcac ctgggtgctc tggctggaga ggaaaagcca gttccctacg
     4141 aggagtgttc ccaatgcttt gtaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4201 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     4261 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     4321 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcgggga
//
