LOCUS       sequence-474037-        5528 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sall4-Fkbp-2xHA targeting vector with Blasto selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5528)
  TITLE     Hendrich lab plasmids batch1
REFERENCE   2  (bases 1 to 5528)
  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: pSall4_GGSG_Fkbp_2xHA_BB
FEATURES             Location/Qualifiers
     source          1..5528
                     /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             2699..3019
                     /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             3026..3052
                     /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             3026..3052
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3056..3082
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    3111..3142
                     /label=rox
                     /bound_moiety="Dre recombinase"
     CDS             3683..4081
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4204..4338
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4241..4260)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4295..4314
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    4354..4385
                     /label=rox
                     /bound_moiety="Dre recombinase"
     primer_bind     complement(4887..4906)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4888..4906)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4913..4930)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4913..4929)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4922..4944)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(5070..5525)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5207..5228)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5419..5438
                     /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 tgacccaaca ataactccgc ccgccgggga aggaagctgg ccatagagaa
     2281 ccccatggtc gcgctgagtg ctgagggaaa gagagcgccc gaggtgtttt ccaaggagct
     2341 cctgtccccc gcggtgagtg tggaccccgc ctcctggaac cagtacacca gcgtcctgaa
     2401 tgggggtctg gccatgaaga ccaacgagat ctccgtgatc cagagcggag gcatccccac
     2461 gctgcctgtg tcgctggggg ccagctctgt ggtgagcaat ggcacgattt ccaagcttga
     2521 cggctctcag accggtgtga gcatgcccat gagcgggaac ggagaaaagc tcgctgttcc
     2581 cgacggcatg gccaaacacc agttccctca cttcctggag gaaaataaga ttgctgtcag
     2641 cgtctctggt ggcggtggct ctggcggagg tgggtccggt ggcggcggaa gcgccggtgg
     2701 agtgcaggtg gaaaccatct ccccaggaga cgggcgcacc ttccccaagc gcggccagac
     2761 ctgcgtggtg cactacaccg ggatgcttga agatggaaag aaagttgatt cctcccggga
     2821 cagaaacaag ccctttaagt ttatgctagg caagcaggag gtgatccgag gctgggaaga
     2881 aggggttgcc cagatgagtg tgggtcagag agccaaactg actatatctc cagattatgc
     2941 ctatggtgcc actgggcacc caggcatcat cccaccacat gccactctcg tcttcgatgt
     3001 ggagcttcta aaactggaag gcggctaccc ctacgacgtg cccgactacg ccggctatcc
     3061 gtatgatgtc ccggactatg caggctatgt cggttaccga taaacgatcg taactttaaa
     3121 taatgccaat tatttaaagt tacatatggc cttctgcagg agcgtacaga acccagggcc
     3181 ctggcacccg tgcagaccct ggcccacccc acctgggcgc tcagtgccca agagatgtcc
     3241 acacctagga tgtcccgcgg tgggtggggg gcccgagaga cgggcaggcc gggggcaggc
     3301 ctggccatgc ggggccgaac cgggcactgc ccagcgtggg gcgcgggggc cacggcgcgc
     3361 gcccccagcc cccgggccca gcaccccaag gcggccaacg ccaaaactct ccctcctcct
     3421 cttcctcaat ctcgctctcg ctcttttttt ttttcgcaaa aggaggggag agggggtaaa
     3481 aaaatgctgc actgtgcggc gaagccggtg agtgagcggc gcggggccaa tcagcgtgcg
     3541 ccgttccgaa agttgccttt tatggctcga gcggccgcgg cggcgcccta taaaacccag
     3601 cggcgcgacg cgccaccacc gccgagaccg cgtccgcccc gcgagcacag agcctcgcct
     3661 ttgccgatcc atcgatggat ccatggccaa gcctttgtct caagaagaat ccaccctcat
     3721 tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca gcgtcgccag
     3781 cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc attttactgg
     3841 gggaccttgc gcagaactcg tggtgctggg cactgctgct gctgcggcag ctggcaacct
     3901 gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct gcggacggtg
     3961 ccgacaggtt cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg
     4021 acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt gggagggcta
     4081 agctctagat aagtaatgat cataatcagc catatcacat ctgtagaggt tttacttgct
     4141 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     4201 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4261 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4321 tcttatcatg tctggatcca tttactgatc acttaacttt aaataatgcc aattatttaa
     4381 agttaggcgc ggggggagtc ttctgcaggg agattctaga atctgcttta ttctttagtt
     4441 tttttttttt ttttttctta acccacctcc tttttttatt ttcttcctct tcctgatatg
     4501 caagtgatgt ggcttgtgac catagccttg ggcaatccta caatcacgag agttgctatg
     4561 ctgctttcca aaaaaaagat gaaaaatgga ggaaaagtga aaaacaacaa aaacaaacca
     4621 aaaaaaataa tgataaaaag ccagactttt tatttttgtt tttaaaccat tttggaaaga
     4681 agcgggtttt gaaaaatagt tctgttaatt tgggacaaat tatattctag aagctttgtg
     4741 tgacctcatg gctgtttcca aagactgtca tcaacctgtt tcgaagtgaa acagttggga
     4801 ctcattggac cacagtggaa gtgacaactc cttaacccag tcctggggtg agggtgtgct
     4861 gtccttctgc gccgatctcc aattcgccct atagtgagtc gtattacaat tcactggccg
     4921 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     4981 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     5041 aacagttgcg cagcctgaat ggcgaatgga aattgtaagc gttaatattt tgttaaaatt
     5101 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     5161 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     5221 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     5281 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     5341 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     5401 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     5461 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     5521 cgcgtcag
//
