LOCUS       sequence-349358-        4442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human PANK2 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: PANK2_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4442
                     /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             2464..3351
                     /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(3809..3825)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3809..3825)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3822..3844)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3833..3849
                     /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(3857..3863,3864..3881,3882..3887))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3902..3923
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4040..4057)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(4211..4442,1..357))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4291..4310)
                     /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 agtgaattcg agctcggtac ctcgcgaatg catctagacc tctgctgctt ctcacctcat
     2041 tccccagaaa cagtcaggtt ctgtctctta ccagaggtaa ctgagctgct gctggcctca
     2101 tagctcctgt cgccctcact gcgggtttgg ttttcctctt atcacagtgg tctgaagtgc
     2161 ttggatagtc taggacatcc agccccctcc ccagacaggg actccttagg gcgtctctgc
     2221 tcccaggctg gccttggccc ttctggggtg ctcaggaaat gggtatgctg tgtgtgggca
     2281 gtggttggct ttaacactag tcatcatgtg taaggtgatt tgaataagtc taaggaaaat
     2341 tgcacttatt tttggtgtat tttctttcag ggttattttg gagctgttgg agcactcctt
     2401 gagctgttga agatcccggg tggaggaggt tctggaggcg gtggaagtgg tggcggaggt
     2461 agcgccgaga tcggcaccgg ctttcccttc gatcctcact acgtggaagt gctgggcgag
     2521 agaatgcact atgtggacgt gggccccaga gatggaaccc ctgtgctgtt tctgcacggc
     2581 aaccctacca gcagctacgt gtggcggaac atcatccctc acgtggcccc tacacacaga
     2641 tgtatcgccc ctgacctgat cggcatgggc aagagcgata agcccgacct gggctacttc
     2701 ttcgacgacc acgtgcggtt catggacgcc tttattgagg ccctcggcct ggaagaggtg
     2761 gtgctggtta ttcacgattg gggcagcgcc ctgggattcc actgggccaa gagaaacccc
     2821 gagagagtga agggaatcgc cttcatggaa ttcatcaggc ccattcctac ctgggacgag
     2881 tggcctgagt tcgccagaga gacattccag gccttcagaa ccaccgacgt gggcagaaag
     2941 ctgatcatcg accagaacgt gttcatcgag ggcaccctgc ctatgggagt cgtcagacca
     3001 ctgaccgagg tggaaatgga ccactacaga gagccctttc tgaaccccgt ggaccgggaa
     3061 cctctttgga gattccccaa cgagctgccc attgccggcg agcctgctaa tattgtggcc
     3121 ctggtggaag agtacatgga ctggctgcat cagagccccg tgcctaagct gctgttttgg
     3181 ggaacacccg gcgtgctgat tcctcctgct gaagctgcca gactggccaa gagcctgcct
     3241 aattgcaagg ccgtggatat cggccctggc ctgaatctgc tgcaagagga caaccccgat
     3301 ctgatcggct ccgagattgc cagatggctg agcaccctgg aaatcagcgg ataatcatta
     3361 cctggggagg ggttcctgaa accttccaca atgggatctg tggactttca tttttttaag
     3421 agacttactc aatttcatga ctgtactacc tgaaacaaag tgagaaagga caggtgtatt
     3481 tttctaagtc atcaagataa atccttaaga attcagtcta aattagcaac caggaaggaa
     3541 aaatatatta aaaacaacaa aaaagtggca catgtccagg cagtgtgagg atttgctgta
     3601 tataagttgc ctgctttgta tttttgaaat ctctgcatca ctcattggaa gtgcttctga
     3661 agagagctgc tctgtgttca gttgactggt tttgtgtcct gtttgaactt gctgaatgta
     3721 aggcaggcta ctatgcgtta taatctaatc acaaggatcc cgggcccgtc gactgcagag
     3781 gcctgcatgc aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     3841 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     3901 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3961 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4021 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4081 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4141 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4201 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4261 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4321 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4381 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4441 tc
//
