LOCUS       sequence-349357-        4442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human ANG 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: ANG_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    12..33
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(150..167)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(321..909)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(401..420)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1080..1871,1872..1940))
                     /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     1703..1722
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1941..2045)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2113..2131
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2169..2191)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2291..2310
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2504..2526
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2518..2535
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2519..2535
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3016..3903
                     /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(4361..4377)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4361..4377)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4374..4396)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4385..4401
                     /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(4409..4415,4416..4433,4434..4439))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
       61 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
      121 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
      181 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
      241 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      301 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      361 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      421 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
      481 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
      541 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
      601 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
      661 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
      721 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
      781 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
      841 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
      901 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
      961 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     1021 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     1081 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     1141 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     1201 agtgctgcaa tgataccgcg agatccacgc tcaccggctc cagatttatc agcaataaac
     1261 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     1321 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     1381 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     1441 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     1501 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     1561 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     1621 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     1681 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     1741 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     1801 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     1861 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     1921 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     1981 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     2041 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     2101 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     2161 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     2221 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     2281 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     2341 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc
     2401 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     2461 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     2521 aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa tgcatctaga gggtctgggt
     2581 ctgaccccac cgaccctggc tcaggataac tccaggtaca cacacttcct gacccagcac
     2641 tatgatgcca aaccacaggg ccgggatgac agatactgtg aaagcatcat gaggagacgg
     2701 ggcctgacct caccctgcaa agacatcaac acatttattc atggcaacaa gcgcagcatc
     2761 aaggccatct gtgaaaacaa gaatggaaac cctcacagag aaaacctaag aataagcaag
     2821 tcttctttcc aggtcaccac ttgcaagcta catggaggtt ccccctggcc tccatgccag
     2881 taccgagcca cagcggggtt cagaaacgtt gttgttgctt gtgaaaatgg cttacctgtc
     2941 cacttggatc agtcaatttt ccgtcgtccg ggtggaggag gttctggagg cggtggaagt
     3001 ggtggcggag gtagcgccga gatcggcacc ggctttccct tcgatcctca ctacgtggaa
     3061 gtgctgggcg agagaatgca ctatgtggac gtgggcccca gagatggaac ccctgtgctg
     3121 tttctgcacg gcaaccctac cagcagctac gtgtggcgga acatcatccc tcacgtggcc
     3181 cctacacaca gatgtatcgc ccctgacctg atcggcatgg gcaagagcga taagcccgac
     3241 ctgggctact tcttcgacga ccacgtgcgg ttcatggacg cctttattga ggccctcggc
     3301 ctggaagagg tggtgctggt tattcacgat tggggcagcg ccctgggatt ccactgggcc
     3361 aagagaaacc ccgagagagt gaagggaatc gccttcatgg aattcatcag gcccattcct
     3421 acctgggacg agtggcctga gttcgccaga gagacattcc aggccttcag aaccaccgac
     3481 gtgggcagaa agctgatcat cgaccagaac gtgttcatcg agggcaccct gcctatggga
     3541 gtcgtcagac cactgaccga ggtggaaatg gaccactaca gagagccctt tctgaacccc
     3601 gtggaccggg aacctctttg gagattcccc aacgagctgc ccattgccgg cgagcctgct
     3661 aatattgtgg ccctggtgga agagtacatg gactggctgc atcagagccc cgtgcctaag
     3721 ctgctgtttt ggggaacacc cggcgtgctg attcctcctg ctgaagctgc cagactggcc
     3781 aagagcctgc ctaattgcaa ggccgtggat atcggccctg gcctgaatct gctgcaagag
     3841 gacaaccccg atctgatcgg ctccgagatt gccagatggc tgagcaccct ggaaatcagc
     3901 ggataaccag cgggcccctg gtcaagtgct ggctctgctg tccttgcctt ccatttcccc
     3961 tctgcaccca gaacagtggt ggcaacattc attgccaagg gcccaaagaa agagctacct
     4021 ggaccttttg ttttctgttt gacaacatgt ttaataaata aaaatgtctt gatatcagta
     4081 agaatcagag tcttctcact gattctgggc atattgatct ttcccccatt ttctctactt
     4141 ggctgctccc tgagaggact gcataggata gaaatgcctt tttcttttct tttcgttttt
     4201 tttttttttt tttttgagat ggagtctcac tctgtcgccc aggcttaagt gcaatggcac
     4261 aatctcggct cactgcaacc tctctctcct gggttcaagt gattctggat cccgggcccg
     4321 tcgactgcag aggcctgcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4381 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     4441 cc
//
