LOCUS       sequence-474319-        6178 bp    DNA     circular SYN 03-JUN-2025
DEFINITION  Sox2-Fkbp-2xHA targeting vector with Blasto selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6178)
  TITLE     Hendrich lab plasmids batch1
REFERENCE   2  (bases 1 to 6178)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 3, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pSox2_GGSG_Fkbp_2xHA_BB
FEATURES             Location/Qualifiers
     source          1..6178
                     /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             3145..3465
                     /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             3472..3498
                     /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             3472..3498
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3502..3528
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4129..4527
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4650..4784
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4687..4706)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4741..4760
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5537..5556)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5538..5556)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5563..5580)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5563..5579)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5572..5594)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(5720..6175)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5857..5878)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6069..6088
                     /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 tgacccagaa gaacagcccg gaccgcgtca agaggcccat gaacgccttc
     2281 atggtatggt cccgggggca gcggcgtaag atggcccagg agaaccccaa gatgcacaac
     2341 tcggagatca gcaagcgcct gggcgcggag tggaaacttt tgtccgagac cgagaagcgg
     2401 ccgttcatcg acgaggccaa gcggctgcgc gctctgcaca tgaaggagca cccggattat
     2461 aaataccggc cgcggcggaa aaccaagacg ctcatgaaga aggataagta cacgcttccc
     2521 ggaggcttgc tggcccccgg cgggaacagc atggcgagcg gggttggggt gggcgccggc
     2581 ctgggtgcgg gcgtgaacca gcgcatggac agctacgcgc acatgaacgg ctggagcaac
     2641 ggcagctaca gcatgatgca ggagcagctg ggctacccgc agcacccggg cctcaacgct
     2701 cacggcgcgg cacagatgca accgatgcac cgctacgacg tcagcgccct gcagtacaac
     2761 tccatgacca gctcgcagac ctacatgaac ggctcgccca cctacagcat gtcctactcg
     2821 cagcagggca cccccggtat ggcgctgggc tccatgggct ctgtggtcaa gtccgaggcc
     2881 agctccagcc cccccgtggt tacctcttcc tcccactcca gggcgccctg ccaggccggg
     2941 gacctccggg acatgatcag catgtacctc cccggcgccg aggtgccgga gcccgctgcg
     3001 cccagtagac tgcacatggc ccagcactac cagagcggcc cggtgcccgg cacggccatt
     3061 aacggcacac tgcccctgtc gcacatgggc tctggtggcg gtggctctgg cggaggtggg
     3121 tccggtggcg gcggaagcgc cggtggagtg caggtggaaa ccatctcccc aggagacggg
     3181 cgcaccttcc ccaagcgcgg ccagacctgc gtggtgcact acaccgggat gcttgaagat
     3241 ggaaagaaag ttgattcctc ccgggacaga aacaagccct ttaagtttat gctaggcaag
     3301 caggaggtga tccgaggctg ggaagaaggg gttgcccaga tgagtgtggg tcagagagcc
     3361 aaactgacta tatctccaga ttatgcctat ggtgccactg ggcacccagg catcatccca
     3421 ccacatgcca ctctcgtctt cgatgtggag cttctaaaac tggaaggcgg ctacccctac
     3481 gacgtgcccg actacgccgg ctatccgtat gatgtcccgg actatgcagg ctatgtcggt
     3541 taccgataaa cgatcgtaac tttaaataat gccaattatt taaagttaca tatggccttc
     3601 tgcaggagcg tacagaaccc agggccctgg cacccgtgca gaccctggcc caccccacct
     3661 gggcgctcag tgcccaagag atgtccacac ctaggatgtc ccgcggtggg tggggggccc
     3721 gagagacggg caggccgggg gcaggcctgg ccatgcgggg ccgaaccggg cactgcccag
     3781 cgtggggcgc gggggccacg gcgcgcgccc ccagcccccg ggcccagcac cccaaggcgg
     3841 ccaacgccaa aactctccct cctcctcttc ctcaatctcg ctctcgctct tttttttttt
     3901 cgcaaaagga ggggagaggg ggtaaaaaaa tgctgcactg tgcggcgaag ccggtgagtg
     3961 agcggcgcgg ggccaatcag cgtgcgccgt tccgaaagtt gccttttatg gctcgagcgg
     4021 ccgcggcggc gccctataaa acccagcggc gcgacgcgcc accaccgccg agaccgcgtc
     4081 cgccccgcga gcacagagcc tcgcctttgc cgatccatcg atggatccat ggccaagcct
     4141 ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc
     4201 atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact
     4261 ggtgtcaatg tatatcattt tactggggga ccttgcgcag aactcgtggt gctgggcact
     4321 gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg
     4381 ggcatcttga gcccctgcgg acggtgccga caggttcttc tcgatctgca tcctgggatc
     4441 aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg
     4501 ccctctggtt atgtgtggga gggctaagct ctagataagt aatgatcata atcagccata
     4561 tcacatctgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     4621 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     4681 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4741 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccattta ctgatcactt
     4801 aactttaaat aatgccaatt atttaaagtt aggcgctgag ggctggactg cgaactggag
     4861 aaggggagag attttcaaag agatacaagg gaattgggag gggtgcaaaa agaggagagt
     4921 aggaaaaatc tgataatgct caaaaggaaa aaaaatctcc gcagcgaaac gacagctgcg
     4981 gaaaaaaacc accaatccca tccaaattaa cgcaaaaacc gtgatgccga ctagaaaact
     5041 tttatgagag atcttgggac ttctttttgg gggactattt ttgtacagag aaaacctgag
     5101 ggcggcgggg agggcggggg aatcggacca tgtatagatc tggaggaaaa aaactacgca
     5161 aaactttttt ttaaagttct agtggtacgt taggcgcttc gcagggagtt cgcaaaagtc
     5221 tttaccagta atatttagag ctagactccg ggcgatgaaa aaaaagtttt aatatttgca
     5281 agcaactttt gtacagtatt tatcgagata aacatggcaa tcaaatgtcc attgtttata
     5341 agctgagaat ttgccaatat ttttcgagga aagggttctt gctgggtttt gattctgcag
     5401 cttaaattta ggaccgttac aaacaaggaa ggagtttatt cggatttgaa cattttagtt
     5461 ttaaaattgt acaaaaggaa aacatgagag caagtactgg caagaccgtt ttcgtggtcc
     5521 gccgatctcc aattcgccct atagtgagtc gtattacaat tcactggccg tcgttttaca
     5581 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     5641 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     5701 cagcctgaat ggcgaatgga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     5761 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     5821 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     5881 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     5941 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     6001 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     6061 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     6121 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcag
//
