LOCUS       sequence-349360-        4442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human PFN1 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: PFN1_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(301..317)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(301..317)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(314..336)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    325..341
                     /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(349..355,356..373,374..379))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    394..415
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(532..549)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(703..1291)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(783..802)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1462..2253,2254..2322))
                     /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     2085..2104
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2323..2427)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2495..2513
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2551..2573)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2673..2692
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2886..2908
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2900..2917
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2901..2917
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3398..4285
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="AEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
ORIGIN
        1 cacctccccc tacccatatc cctcccgtgt gtggttggaa aacttttgtt ttttggggtt
       61 ttttttttct gaataaaaaa gattctacta acaaggtgct gtgtgatttg agcttgggtg
      121 tcagaatggg ggataggcac tgtgacttgc ctcctttatc gtggcttggg agcagggagg
      181 ttgaggtact acaaggtgca ggaatttgag cttctggcca ggtggccctg cccctccaca
      241 gtggcaggat cccgggcccg tcgactgcag aggcctgcat gcaagcttgg cgtaatcatg
      301 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
      361 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
      421 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
      481 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
      541 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
      601 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
      661 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
      721 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
      781 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
      841 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
      901 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
      961 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     1021 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     1081 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     1141 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     1201 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     1261 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     1321 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     1381 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     1441 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     1501 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     1561 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagatccac gctcaccggc
     1621 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     1681 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     1741 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     1801 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     1861 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     1921 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     1981 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     2041 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     2101 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     2161 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     2221 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     2281 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     2341 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     2401 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     2461 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     2521 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     2581 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     2641 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     2701 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     2761 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     2821 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     2881 gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttcgagctcg gtacctcgcg
     2941 aatgcatcta gaaacctgga ggtggcatgc acatccacag ccctttctag cctgaacaca
     3001 tgaatcaaca gtctatgagc ctttgacccc taagttttta gtctgttcct ctccagcgtt
     3061 taagcttgat tttcctcttt tgaaaagcct tattgtttca gggtgactga cagcctgcct
     3121 gtgtgtggtg ggggggcggg ttgggagaga tgaggttggg ttacagcccc cagggctgga
     3181 cagctgctga acagctggca gggggtggaa ttgtgacacc tggatcctag cctcctttct
     3241 ctttcctcct cctccagcgc tagtcctgct gatgggcaaa gaaggtgtcc acggtggttt
     3301 gatcaacaag aaatgttatg aaatggcctc ccaccttcgg cgttcccagt acggtggagg
     3361 aggttctgga ggcggtggaa gtggtggcgg aggtagcgcc gagatcggca ccggctttcc
     3421 cttcgatcct cactacgtgg aagtgctggg cgagagaatg cactatgtgg acgtgggccc
     3481 cagagatgga acccctgtgc tgtttctgca cggcaaccct accagcagct acgtgtggcg
     3541 gaacatcatc cctcacgtgg cccctacaca cagatgtatc gcccctgacc tgatcggcat
     3601 gggcaagagc gataagcccg acctgggcta cttcttcgac gaccacgtgc ggttcatgga
     3661 cgcctttatt gaggccctcg gcctggaaga ggtggtgctg gttattcacg attggggcag
     3721 cgccctggga ttccactggg ccaagagaaa ccccgagaga gtgaagggaa tcgccttcat
     3781 ggaattcatc aggcccattc ctacctggga cgagtggcct gagttcgcca gagagacatt
     3841 ccaggccttc agaaccaccg acgtgggcag aaagctgatc atcgaccaga acgtgttcat
     3901 cgagggcacc ctgcctatgg gagtcgtcag accactgacc gaggtggaaa tggaccacta
     3961 cagagagccc tttctgaacc ccgtggaccg ggaacctctt tggagattcc ccaacgagct
     4021 gcccattgcc ggcgagcctg ctaatattgt ggccctggtg gaagagtaca tggactggct
     4081 gcatcagagc cccgtgccta agctgctgtt ttggggaaca cccggcgtgc tgattcctcc
     4141 tgctgaagct gccagactgg ccaagagcct gcctaattgc aaggccgtgg atatcggccc
     4201 tggcctgaat ctgctgcaag aggacaaccc cgatctgatc ggctccgaga ttgccagatg
     4261 gctgagcacc ctggaaatca gcggataacc tcgtctgtcc cttccccttc accgctcccc
     4321 acagctttgc acccctttcc tccccataca cacacaaacc attttatttt ttgggccatt
     4381 accccatacc ccttattgct gccaaaacca catgggctgg gggccagggc tggatggaca
     4441 ga
//
