LOCUS       sequence-473993-        5456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Targeting construct: Chd8-FKBP-HA2 with Roxed Blasto cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5456)
  TITLE     Hendrich lab plasmids batch2
REFERENCE   2  (bases 1 to 5456)
  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: pChd8_GGSG_Fkbp_2xHA_BB
FEATURES             Location/Qualifiers
     source          1..5456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(129..197,198..989)
                     /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     complement(347..366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1649..1668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1902..1919
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2072..2077,2078..2095,2096..2102)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /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)."
     primer_bind     2115..2137
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2134..2150
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2171..2189
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             2466..2483
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2493..2513
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             2790..3110
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=FKBP-V
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             3117..3143
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             3117..3143
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3147..3173
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    3202..3233
                     /label=rox
                     /bound_moiety="Dre recombinase"
     CDS             3774..4172
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4295..4429
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4332..4351)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4386..4405
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    4445..4476
                     /label=rox
                     /bound_moiety="Dre recombinase"
     primer_bind     complement(4815..4834)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4816..4834)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4841..4858)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4841..4857)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4850..4872)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(4998..5453)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5135..5156)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5347..5366
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     2161 ccaagctcga aattaaccct cactaaaggg aacaaaagct ggtaccgggc cccccctcga
     2221 ggtcgaagct tgacagcaag aaacgggtcc gtaggacaag accagacctt tctaagatga
     2281 tggccctgat gcagggtgga agcactgggt ccctgtctct gcataacacc tttcaacaca
     2341 gcagtagtaa cctacagtct gtgtcgtctc tgggtcacag cagtaccact tctgcatctc
     2401 tgccttttat gccgtttgtg atgggtgcag cagcaccccc tcatgtagac tccagcacca
     2461 tgcttcatca tcaccaccac catccccacc cccaccatca ccaccatcac catccaggct
     2521 tgagaaccac tggctaccct tcttcaccag ctactaccac ctctggtact gccttgcggt
     2581 taccaacact gcaacctgag gacgatgatg aagaggagga tgaagaagat gatgatttat
     2641 ctcagggcta tgacagctca gaacgggact tctcactcat tgatgaccct atgatgccag
     2701 ccaactcaga ctccagtgaa gatgctgacg acggctctgg tggcggtggc tctggcggag
     2761 gtgggtccgg tggcggcgga agcgccggtg gagtgcaggt ggaaaccatc tccccaggag
     2821 acgggcgcac cttccccaag cgcggccaga cctgcgtggt gcactacacc gggatgcttg
     2881 aagatggaaa gaaagttgat tcctcccggg acagaaacaa gccctttaag tttatgctag
     2941 gcaagcagga ggtgatccga ggctgggaag aaggggttgc ccagatgagt gtgggtcaga
     3001 gagccaaact gactatatct ccagattatg cctatggtgc cactgggcac ccaggcatca
     3061 tcccaccaca tgccactctc gtcttcgatg tggagcttct aaaactggaa ggcggctacc
     3121 cctacgacgt gcccgactac gccggctatc cgtatgatgt cccggactat gcaggctatg
     3181 tcggttaccg ataaacgatc gtaactttaa ataatgccaa ttatttaaag ttacatatgg
     3241 ccttctgcag gagcgtacag aacccagggc cctggcaccc gtgcagaccc tggcccaccc
     3301 cacctgggcg ctcagtgccc aagagatgtc cacacctagg atgtcccgcg gtgggtgggg
     3361 ggcccgagag acgggcaggc cgggggcagg cctggccatg cggggccgaa ccgggcactg
     3421 cccagcgtgg ggcgcggggg ccacggcgcg cgcccccagc ccccgggccc agcaccccaa
     3481 ggcggccaac gccaaaactc tccctcctcc tcttcctcaa tctcgctctc gctctttttt
     3541 tttttcgcaa aaggagggga gagggggtaa aaaaatgctg cactgtgcgg cgaagccggt
     3601 gagtgagcgg cgcggggcca atcagcgtgc gccgttccga aagttgcctt ttatggctcg
     3661 agcggccgcg gcggcgccct ataaaaccca gcggcgcgac gcgccaccac cgccgagacc
     3721 gcgtccgccc cgcgagcaca gagcctcgcc tttgccgatc catcgatgga tccatggcca
     3781 agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca
     3841 tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct
     3901 tcactggtgt caatgtatat cattttactg ggggaccttg cgcagaactc gtggtgctgg
     3961 gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga
     4021 acaggggcat cttgagcccc tgcggacggt gccgacaggt tcttctcgat ctgcatcctg
     4081 ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat
     4141 tgctgccctc tggttatgtg tgggagggct aagctctaga taagtaatga tcataatcag
     4201 ccatatcaca tctgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     4261 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     4321 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4381 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc atttactgat
     4441 cacttaactt taaataatgc caattattta aagttaggcg agccccatat agacccgtga
     4501 tgggtggctg ctggaatccc ttactctggc ccttggactt tggaaatgtg ggaggggcag
     4561 ggaacaatat ggggaaaccc aagacttcag gaggtgaaaa gaaaaagaaa aaaacaaaaa
     4621 acaaaaaaaa aaaaaaaaaa acttgtaccc aattgctccc aattttgaga ggattgggtg
     4681 ggcaggggaa ctcctaaaaa aaaataataa taaatgacca cttcctcgtt tctggggaag
     4741 caaaggagct tagagcagct ggtgtgctca cccctcccca gacgcctcca ttaaccacgc
     4801 cgatctccaa ttcgccctat agtgagtcgt attacaattc actggccgtc gttttacaac
     4861 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     4921 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     4981 gcctgaatgg cgaatggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     5041 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     5101 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     5161 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     5221 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     5281 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     5341 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     5401 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcag
//
