LOCUS       sequence-473984-        5726 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Adnp-Fkbp-2xHA targeting vector with Blasto selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5726)
  TITLE     Hendrich lab plasmids batch3
REFERENCE   2  (bases 1 to 5726)
  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: pAdnp_GGSG_Fkbp_2xHA_BB
FEATURES             Location/Qualifiers
     source          1..5726
                     /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             2866..3186
                     /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             3193..3219
                     /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             3193..3219
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3223..3249
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    3278..3309
                     /label=rox
                     /bound_moiety="Dre recombinase"
     CDS             3850..4248
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4371..4505
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4408..4427)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4462..4481
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    4521..4552
                     /label=rox
                     /bound_moiety="Dre recombinase"
     primer_bind     complement(5085..5104)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5086..5104)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5111..5128)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5111..5127)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5120..5142)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(5268..5723)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5405..5426)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5617..5636
                     /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 tgactccgga aggtgctttg gaatctgaga agctagacca aaaagaggag
     2281 gaggaggagg aggaggatgg ttcaaaatat gaaactatcc atttgactga ggaaccagcc
     2341 aaattaatgc atgatgcctc tgatagtgag gtagaccaag atgatgtagt tgagtggaaa
     2401 gatggtgctt caccatctga gagtgggcct ggttcccaac aaatctcaga ctttgaggat
     2461 aatacatgtg aaatgaaacc aggaacctgg tctgatgagt cttcccagag tgaagatgca
     2521 aggagcagta agccagctgc caaaaaaaag gctacagtgc aagatgacac agagcagtta
     2581 aaatggaaga atagttccta tggaaaagtt gaagggtttt ggtccaagga ccagtcacag
     2641 tgggaaaatg catctgagaa tgcagagcgc ttaccaaacc cacagattga gtggcagaat
     2701 agcacaattg acagtgagga cggggagcag tttgacagca tgactgacgg agttgctgat
     2761 cccatgcatg gcagcttaac tggagtgaag ctgagcagcc agcaagccgg ctctggtggc
     2821 ggtggctctg gcggaggtgg gtccggtggc ggcggaagcg ccggtggagt gcaggtggaa
     2881 accatctccc caggagacgg gcgcaccttc cccaagcgcg gccagacctg cgtggtgcac
     2941 tacaccggga tgcttgaaga tggaaagaaa gttgattcct cccgggacag aaacaagccc
     3001 tttaagttta tgctaggcaa gcaggaggtg atccgaggct gggaagaagg ggttgcccag
     3061 atgagtgtgg gtcagagagc caaactgact atatctccag attatgccta tggtgccact
     3121 gggcacccag gcatcatccc accacatgcc actctcgtct tcgatgtgga gcttctaaaa
     3181 ctggaaggcg gctaccccta cgacgtgccc gactacgccg gctatccgta tgatgtcccg
     3241 gactatgcag gctatgtcgg ttaccgataa acgatcgtaa ctttaaataa tgccaattat
     3301 ttaaagttac atatggcctt ctgcaggagc gtacagaacc cagggccctg gcacccgtgc
     3361 agaccctggc ccaccccacc tgggcgctca gtgcccaaga gatgtccaca cctaggatgt
     3421 cccgcggtgg gtggggggcc cgagagacgg gcaggccggg ggcaggcctg gccatgcggg
     3481 gccgaaccgg gcactgccca gcgtggggcg cgggggccac ggcgcgcgcc cccagccccc
     3541 gggcccagca ccccaaggcg gccaacgcca aaactctccc tcctcctctt cctcaatctc
     3601 gctctcgctc tttttttttt tcgcaaaagg aggggagagg gggtaaaaaa atgctgcact
     3661 gtgcggcgaa gccggtgagt gagcggcgcg gggccaatca gcgtgcgccg ttccgaaagt
     3721 tgccttttat ggctcgagcg gccgcggcgg cgccctataa aacccagcgg cgcgacgcgc
     3781 caccaccgcc gagaccgcgt ccgccccgcg agcacagagc ctcgcctttg ccgatccatc
     3841 gatggatcca tggccaagcc tttgtctcaa gaagaatcca ccctcattga aagagcaacg
     3901 gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc agctctctct
     3961 agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg accttgcgca
     4021 gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc
     4081 gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg acaggttctt
     4141 ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca gccgacggca
     4201 gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaagc tctagataag
     4261 taatgatcat aatcagccat atcacatctg tagaggtttt acttgcttta aaaaacctcc
     4321 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     4381 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4441 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     4501 ggatccattt actgatcact taactttaaa taatgccaat tatttaaagt taggcgctat
     4561 gcatatgggc cgtgttgcat cctggacttc tgctctcctt ccagtctgac tgcaaagctg
     4621 tcttctaact ggcactacct tgcaaggact ggtcagtcag caggctgtgg ggatgtgtga
     4681 ccactgtagt ctcagtggtt atttccaagt ctatgataga tgactggttg atctttgttc
     4741 agactcttct ctggtagact cggtgatgaa gtgtgaaaaa ccagtaagct ggtagctcat
     4801 gaatgcacac agggagaagt agttccttac ctacctgcct tctcagcatt tcttcccctc
     4861 agatgtttgg ttggatgtcc tgggaaagcc ttatcctgat tcacatagta gtgcagggcg
     4921 ggcatacaca ctaccagtca aatgtgtcta gtagacttgg gaacatggac cgttttttca
     4981 tgtatcttct gaatagttga aatgtatatt tgtacagtct tttagaccta aagtgatgct
     5041 tatgacactg ttactgtgtg cccgtcacgc cgatctccaa ttcgccctat agtgagtcgt
     5101 attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     5161 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     5221 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggaaa ttgtaagcgt
     5281 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     5341 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     5401 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     5461 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     5521 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     5581 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     5641 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     5701 taatgcgccg ctacagggcg cgtcag
//
