LOCUS       sequence-290701-        5715 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsPTB2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5715)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 5715)
  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: pT7-V5-SBP-C1-HsPTB2_AA
FEATURES             Location/Qualifiers
     source          1..5715
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     613..632
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        613..631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      659..668
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             668..709
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             731..844
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWGGGHVVEGLAGELEQLRARLEHHPQGQREP"
     polyA_signal    2503..2624
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2540..2559)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2594..2613
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2631..3086)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2768..2789)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2980..2999
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3113..3217
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3219..3576
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3219..3410
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3242..3262)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3427..3562
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3611..4405
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3611..4405
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3665..3684)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4275..4294
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4593..4612)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4637..4684
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5013..5601
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5502..5521
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      601 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
      661 caccatgggt aagcctatcc ctaaccctct cctcggtctc gattctacgc ccggaagcgg
      721 tagcggtacg atggacgaga agaccaccgg ctgggggggc ggccacgtgg tggagggcct
      781 ggccggcgag ctggagcagc tgcgggccag gctggagcac caccctcagg gccagcggga
      841 gcccagatct cgagctcaag cttcgaattc catggacgga atcgtcactg aggttgcagt
      901 tggcgtgaag agaggatctg acgaactact ctcaggcagt gttctcagta gtccgaactc
      961 taatatgagc agcatggtag ttacagccaa tggtaatgat agtaaaaaat ttaaaggaga
     1021 agataaaatg gatggtgctc cttctcgtgt acttcatatt cgaaaattac ctggggaagt
     1081 aacagaaact gaagttattg ctttaggctt accttttggt aaggtgacca acatccttat
     1141 gctgaaagga aaaaatcagg catttttgga actagcaacc gaggaagcag ctattactat
     1201 ggttaattac tattctgctg tgacacctca tcttcgtaac caaccaatat atatccagta
     1261 ctcgaatcac aaagaactaa agacagataa tacattaaac caacgtgctc aggcagttct
     1321 tcaagctgtg acagctgtcc agacagcaaa tactcctctt agtggcacca cagttagcga
     1381 gagtgcagtg actccagccc agagtccagt acttagaata attattgaca acatgtacta
     1441 ccctgtaaca cttgatgttc ttcaccaaat attttctaag tttggtgctg tattgaagat
     1501 aatcacattt acaaaaaata accagtttca agctttgctc cagtatggtg atccagtaaa
     1561 tgctcaacaa gcaaaactag ccctagatgg tcagaatatt tataatgcct gctgtaccct
     1621 aaggattgat ttttccaaac ttgtgaattt gaatgtaaaa tacaacaatg ataaaagtag
     1681 ggattatact cgacctgatc ttccatctgg ggatggacaa cctgcattgg acccagctat
     1741 tgctgcagca tttgccaagg agacatccct cttagctgtt ccaggagctc tgagtccttt
     1801 ggccattcca aatgctgctg cagcagctgc tgcagctgct gctggccgag tgggtatgcc
     1861 tggagtctca gctggtggca atacagtcct gttggttagc aatttaaatg aagaggtgtt
     1921 tatggagatg tgcagcgtgt gaagatttta tacaataaga aagacagcgc tctaatacag
     1981 atggctgatg gaaaccaatc acaacttgcc atgaatcatc ttaatggaca gaaaatgtat
     2041 ggaaaaatta ttcgtgttac tctgtctaaa catcagactg tacagctacc tcgagaggga
     2101 cttgatgatc aagggctaac aaaagatttt ggtaattccc cattgcatcg ttttaagaaa
     2161 cctggatcca aaaattttca aaacattttt cctccttctg ccacccttca cctatctaat
     2221 atcccaagag atcacaaaat ggctcttctt cagatggcaa cagtggaaga agctattcag
     2281 gccttgattg atcttcataa ttataacctt ggagaaaacc atcatctgag agtgtctttc
     2341 tccaagtcaa caatttaagg taccgcgggc ccgggatcca ccggatctag ataactgatc
     2401 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     2461 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     2521 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     2581 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag
     2641 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     2701 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     2761 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     2821 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     2881 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     2941 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     3001 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     3061 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     3121 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3181 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     3241 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3301 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3361 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3421 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3481 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     3541 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     3601 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     3661 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     3721 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     3781 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     3841 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     3901 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     3961 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4021 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4081 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     4141 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4201 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4261 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4321 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4381 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     4441 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4501 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4561 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     4621 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     4681 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     4741 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     4801 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     4861 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4921 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4981 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5041 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5101 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     5161 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5221 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5281 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5341 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5401 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5461 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5521 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5581 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5641 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5701 attaccgcca tgcat
//
