LOCUS       sequence-349324-        4442 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Donor vector for endogenous tagging of human SNCB 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 Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: SNCB_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(88..105)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(259..847)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(339..358)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1018..1809,1810..1878))
                     /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     1641..1660
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1879..1983)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2051..2069
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2107..2129)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2229..2248
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2442..2464
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2456..2473
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2457..2473
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2954..3841
                     /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(4299..4315)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4299..4315)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4312..4334)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4323..4339
                     /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(4347..4353,4354..4371,4372..4377))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4392..4413
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
       61 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
      121 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
      181 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
      241 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
      301 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      361 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      421 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
      481 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
      541 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
      601 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
      661 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
      721 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
      781 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
      841 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
      901 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
      961 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1021 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1081 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     1141 tgctgcaatg ataccgcgag atccacgctc accggctcca gatttatcag caataaacca
     1201 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1261 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1321 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1381 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1441 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1501 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1561 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     1621 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     1681 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     1741 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     1801 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     1861 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     1921 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     1981 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     2041 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     2101 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     2161 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     2221 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     2281 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     2341 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     2401 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     2461 aacgacggcc agtgaattcg agctcggtac ctcgcgaatg catctagagt gagggggcag
     2521 cagggctggg cgggactttg ggaatccagg atgattgctg ccgtccctag ctggggtggg
     2581 acatccctgg catgagatgg ttgggccagg gagcttgaca tccctcttct agaacccagg
     2641 ccccaggagg tgggagcagg gaggagaggg cagatagggt ctagcgctgg gccccaggca
     2701 cacctgccca agaattctct gttgtcccca acttccaggc tcctcccctg cccaggaaaa
     2761 ggggaatgtg ctattcctcc ttcagggaga gggttcttgg gccggggctt ggccacttgg
     2821 tctcaaactc ctcctccttt ttgctcttct ctactccacc cccatcccgg tgatgcagga
     2881 ggaatatcag gagtatgagc cagaggcggg tggaggaggt tctggaggcg gtggaagtgg
     2941 tggcggaggt agcgccgaga tcggcaccgg ctttcccttc gatcctcact acgtggaagt
     3001 gctgggcgag agaatgcact atgtggacgt gggccccaga gatggaaccc ctgtgctgtt
     3061 tctgcacggc aaccctacca gcagctacgt gtggcggaac atcatccctc acgtggcccc
     3121 tacacacaga tgtatcgccc ctgacctgat cggcatgggc aagagcgata agcccgacct
     3181 gggctacttc ttcgacgacc acgtgcggtt catggacgcc tttattgagg ccctcggcct
     3241 ggaagaggtg gtgctggtta ttcacgattg gggcagcgcc ctgggattcc actgggccaa
     3301 gagaaacccc gagagagtga agggaatcgc cttcatggaa ttcatcaggc ccattcctac
     3361 ctgggacgag tggcctgagt tcgccagaga gacattccag gccttcagaa ccaccgacgt
     3421 gggcagaaag ctgatcatcg accagaacgt gttcatcgag ggcaccctgc ctatgggagt
     3481 cgtcagacca ctgaccgagg tggaaatgga ccactacaga gagccctttc tgaaccccgt
     3541 ggaccgggaa cctctttgga gattccccaa cgagctgccc attgccggcg agcctgctaa
     3601 tattgtggcc ctggtggaag agtacatgga ctggctgcat cagagccccg tgcctaagct
     3661 gctgttttgg ggaacacccg gcgtgctgat tcctcctgct gaagctgcca gactggccaa
     3721 gagcctgcct aattgcaagg ccgtggatat cggccctggc ctgaatctgc tgcaagagga
     3781 caaccccgat ctgatcggct ccgagattgc cagatggctg agcaccctgg aaatcagcgg
     3841 ataagggccc aggagagccc ccaccagcag cacaattctg tccctgtccc tgccccgccc
     3901 cccagagcca gggctgtcct tagactcctt ctccccaatc acgagatctt ccttccgctc
     3961 tgaggcaacc ccctcggagc ctgtgttagt gtctgtccat ctgtctgtcc tacccgcccg
     4021 cgtccaaccc cggggcatgg acagggccag ggttgcggtc gcggctggga gcctcgcccc
     4081 tccagtgttg cctcctccca tccagcgtct gcgcggatgt agcatgttct atgtgttttt
     4141 aaacgaagat ccgagcgacg gctcctcccc gatccccgac agtggctctc caagcggccc
     4201 ccgggcagcc ccagagcacc ccgcccgact cccattaacc tcgaggatcc cgggcccgtc
     4261 gactgcagag gcctgcatgc aagcttggcg taatcatggt catagctgtt tcctgtgtga
     4321 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4381 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4441 ca
//
