LOCUS       Exported                4442 bp ds-DNA     circular SYN 02-DEC-2021
DEFINITION  Donor vector for endogenous tagging of human ERBB4 at the C-terminus
            with halotag.
ACCESSION   .
VERSION     .
KEYWORDS    ERBB4_Halo_C_allele
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 2, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4442
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(146..734)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(226..245)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(905..1765)
                     /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     1528..1547
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1766..1870)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1938..1956
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1994..2016)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2116..2135
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2329..2351
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2343..2360
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2344..2360
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2841..3728
                     /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(4186..4202)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4186..4202)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4199..4221)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4210..4226
                     /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(4234..4264)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4279..4300
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4417..4434)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
       61 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
      121 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      181 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      241 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      301 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
      361 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
      421 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
      481 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
      541 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
      601 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
      661 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
      721 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
      781 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
      841 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
      901 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
      961 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1021 gccccagtgc tgcaatgata ccgcgagatc cacgctcacc ggctccagat ttatcagcaa
     1081 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1141 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1201 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1261 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     1321 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     1381 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     1441 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     1501 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     1561 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     1621 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     1681 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     1741 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     1801 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     1861 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     1921 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     1981 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     2041 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     2101 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     2161 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     2221 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     2281 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     2341 gttgtaaaac gacggccagt gaattcgagc tcggtacctc gcgaatgcat ctagaaaaaa
     2401 atggagacct tcaagcattg gataatcccg aatatcacaa tgcatccaat ggtccaccca
     2461 aggccgagga tgagtatgtg aatgagccac tgtacctcaa cacctttgcc aacaccttgg
     2521 gaaaagctga gtacctgaag aacaacatac tgtcaatgcc agagaaggcc aagaaagcgt
     2581 ttgacaaccc tgactactgg aaccacagcc tgccacctcg gagcaccctt cagcacccag
     2641 actacctgca ggagtacagc acaaaatatt tttataaaca gaatgggcgg atccggccta
     2701 ttgtggcaga gaatcctgaa tacctctctg agttctccct gaagccaggc actgtgctgc
     2761 cgcctccacc ttacagacac cggaatactg tggtgggtgg aggaggttct ggaggcggtg
     2821 gaagtggtgg cggaggtagc gccgagatcg gcaccggctt tcccttcgat cctcactacg
     2881 tggaagtgct gggcgagaga atgcactatg tggacgtggg ccccagagat ggaacccctg
     2941 tgctgtttct gcacggcaac cctaccagca gctacgtgtg gcggaacatc atccctcacg
     3001 tggcccctac acacagatgt atcgcccctg acctgatcgg catgggcaag agcgataagc
     3061 ccgacctggg ctacttcttc gacgaccacg tgcggttcat ggacgccttt attgaggccc
     3121 tcggcctgga agaggtggtg ctggttattc acgattgggg cagcgccctg ggattccact
     3181 gggccaagag aaaccccgag agagtgaagg gaatcgcctt catggaattc atcaggccca
     3241 ttcctacctg ggacgagtgg cctgagttcg ccagagagac attccaggcc ttcagaacca
     3301 ccgacgtggg cagaaagctg atcatcgacc agaacgtgtt catcgagggc accctgccta
     3361 tgggagtcgt cagaccactg accgaggtgg aaatggacca ctacagagag ccctttctga
     3421 accccgtgga ccgggaacct ctttggagat tccccaacga gctgcccatt gccggcgagc
     3481 ctgctaatat tgtggccctg gtggaagagt acatggactg gctgcatcag agccccgtgc
     3541 ctaagctgct gttttgggga acacccggcg tgctgattcc tcctgctgaa gctgccagac
     3601 tggccaagag cctgcctaat tgcaaggccg tggatatcgg ccctggcctg aatctgctgc
     3661 aagaggacaa ccccgatctg atcggctccg agattgccag atggctgagc accctggaaa
     3721 tcagcggata agctcagttg tggtttttta ggtggagaga cacacctgct ccaatttccc
     3781 cacccccctc tctttctctg gtggtcttcc ttctacccca aggccagtag ttttgacact
     3841 tcccagtgga agatacagag atgcaatgat agttatgtgc ttacctaact tgaacattag
     3901 agggaaagac tgaaagagaa agataggagg aaccacaatg tttcttcatt tctctgcatg
     3961 ggttggtcag gagaatgaaa cagctagaga aggaccagaa aatgtaaggc aatgctgcct
     4021 actatcaaac tagctgtcac tttttttctt tttctttttc tttctttgtt tctttcttcc
     4081 tcttcttttt tttttttttt tttaaagcag atggttgaaa cacccatgct aggatcccgg
     4141 gcccgtcgac tgcagaggcc tgcatgcaag cttggcgtaa tcatggtcat agctgtttcc
     4201 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4261 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4321 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4381 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4441 gg
//
