LOCUS       sequence-449448-        6469 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Gateway entry vector with a clover-tagged phosphatase-inactive Shp2 
            mutant (D425A,C459S).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6469)
  TITLE     Dynamic regulation of integrin beta-1 phosphorylation supports 
            invasive breast cancer progression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6469)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pENTR2b-PTPN11(D425A,C459S)-Clover
FEATURES             Location/Qualifiers
     source          1..6469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      912..921
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          957..1783
                     /label=CMV intron A
                     /note="human cytomegalovirus intron A (Chapman et al., 
                     1991)"
     primer_bind     complement(1898..1922)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(1919..2122)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1948..1968)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2123..2426
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2601..3189)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2681..2700)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3518..3565
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3590..3609
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3797..4591)
                     /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             complement(3797..4591)
                     /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(3908..3927)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4518..4537
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     enhancer        4738..4929
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4886..4906
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4931..5035)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5062..5517
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5149..5168)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5359..5380
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5524..5748
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 cacgccgtcc acgtaccaat tgaacttcac ctctgggtcc tcgtgggaca cgtccaccac
       61 cacgcacgtc acctcagggg tccgggagat catcagggtg tccttaggct taggagggaa
      121 cagaaacacg cttggtccgc ccagcagctc gggagctggg catggagggc acgtgtgggt
      181 cttgtcgcag gacttaggct ccaccttctt gtccaccttg gtgttggaag gcttgtggtt
      241 cacgttgcag atgtaggtct gggtgcccag ggaggaggaa ggcacggtca ccacggagga
      301 cagggagtac aggccggagg actgcagcac ggcagggaag gtgtgcacgc cgctggtcag
      361 ggcgccagag ttccaggaca cggtcacagg ctcagggaag tagtccttca ccaggcagcc
      421 cagagcggcg gtgccgccgg aggtggactt ggaggaaggg gccagaggga acacggaagg
      481 gcccttggtg gaggcggagc tcactgtcac ggtggtgcct gtgccccaca catcgaagta
      541 ccaggaggag ccgtagtagt ccttcctgcc acagtagtac ctggcggtat catcggactg
      601 caggctattc atcttcagga acacctggct cttggagtta tccttggaga tgctcagcct
      661 gctcttcagg gcggaattgt agttggtgcc gccgcctgtc cagatcactc ccagccactc
      721 caggcccttg ccgggaggct gcctcaccca ggagatggcg taggaggtca gggagaagcc
      781 actaacagta caggtgatgc tcaggctctg agatggggcc accaggccag ggccggactc
      841 cttcagctgc acctgtccgg tggtatcagg gagccagagt agcaggagga agagaagctg
      901 cgctggggct tccatggtgg cggcaagctt cccgagacgc gtgtcaagga cggtgactgc
      961 agaaaagacc catggaaagg aacagtctgt tagtctgtca gctattatgt ctggtggcgc
     1021 gcgcggcagc aacgagtact gctcagacta cactgccctc caccgttaac agcaccgcaa
     1081 cgggagttac ctctgactct tatcagaaca caacaactca gctgcctgca tcttcttctg
     1141 ccgctgcctt aagtcttcca aatgcgtcag cggtgcaagc ccgctccccg agctcatttt
     1201 cagacacata ccctaccgcc acggccttgt gcggcacact ggtggtggtg ggcatcgtgc
     1261 tgtgcctaag tctggcctcc actgttagga gcaaggagct gccgagcgac catgagtcgc
     1321 tggaggcatg ggagcagggc tcggatgtag aagctccgcc gctaccggag aagagcccat
     1381 gtccggaaca cgtacccgag attcgcgtgg agatcccacg ttatgtttaa taaaaactgc
     1441 gggcactggg gacggtggtg ttgtatatgt gaatttgtaa ataataaatg agaccccatc
     1501 ctgtaaaaat acagagtccg tgtcagtctc tgaaggacag tgtattggca tatagccaat
     1561 aaagagagtt gtggcaaaga gccatgttat ggattagtaa tggaaagtat cgtcaccaat
     1621 aggggagtgg tcaataatgg tcaataaccc acacctatag gctaagctat accatcacct
     1681 ataacatgag gaagcggggg tgtatagacc ccaagccaaa aacagtatag catgcataag
     1741 aagccaaggg ggtgggccta tagactctat aggcggtact tacgtcactc ttggcacggg
     1801 gaatccgcgt tccaatgcac cgttcccggc cgcggaggct ggatcggtcc cggtgtcttc
     1861 tatggaggtc aaaacagcgt ggatggcgtc tccaggcgat ctgacggttc actaaacgag
     1921 ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg
     1981 agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga
     2041 cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga
     2101 tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc
     2161 aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt
     2221 cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg
     2281 cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta
     2341 tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
     2401 ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc
     2461 ccgtaattga ttactattaa taactaatgc atggcggtaa tacggttatc cacagaatca
     2521 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2581 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2641 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2701 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2761 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2821 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2881 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2941 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3001 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3061 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3121 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3181 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3241 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3301 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaacctga ggctatggca
     3361 gggcctgccg ccccgacgtt ggctgcgagc cctgggcctt cacccgaact tggggggtgg
     3421 ggtggggaaa aggaagaaac gcgggcgtat tggccccaat ggggtctcgg tggggtatcg
     3481 acagagtgcc agccctggga ccgaaccccg cgtttatgaa caaacgaccc aacaccgtgc
     3541 gttttattct gtctttttat tgccgtcata gcgcgggttc cttccggtat tgtctccttc
     3601 cgtgtttcag ttagcctccc cctagggtgg gcgaagaact ccagcatgag atccccgcgc
     3661 tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag
     3721 gcggcggtgg aatcgaaatc tcgtgatggc acgttgggcg tcgcttggtc ggtcatttcg
     3781 aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
     3841 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagtt
     3901 cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc
     3961 ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
     4021 catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg agcctggcga
     4081 acagttcggc tggcgcgagc ccctgatgtt cttcgtccag atcatcctga tcgacaagac
     4141 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
     4201 acgtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
     4261 cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc
     4321 agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg
     4381 ccagccacga tagccgcgct gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg
     4441 tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc
     4501 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag
     4561 aaccagcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat
     4621 cgatctttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca
     4681 gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcggg
     4741 actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
     4801 ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct
     4861 gctggggagc ctggggactt tccacaccct aactgacaca cattccacag ctggttcttt
     4921 ccgcctcagg attcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4981 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5041 ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     5101 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     5161 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     5221 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     5281 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     5341 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     5401 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     5461 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttacg
     5521 cgtccataga gcccaccgca tccccagcat gcctgctatt gtcttcccaa tcctccccct
     5581 tgctgtcctg ccccacccca ccccccagaa tagaatgaca cctactcaga caatgcgatg
     5641 caatttcctc attttattag gaaaggacag tgggagtggc accttccagg gtcaaggaag
     5701 gcacggggga ggggcaaaca acagatggct ggcaactaga aggcacaggg taccagatat
     5761 cataacttaa aactgccaca cgcaaaaaag aaaaccaaag tggtccacaa aacattctcc
     5821 tttccttctg aaggttttac gatgcattgt tatcattaac cagtctttta ctactaaact
     5881 taaatggcca attgaaacaa acagttctga gaccgttctt ccaccactga ttaagagtgg
     5941 ggtggcacgt attagggata atattctcga gtcatttacc cggggacagg gagaggctct
     6001 tctgggtgta gtggttgtgc agggcctcgt gcatcacgga gcaggagaac acgttgccct
     6061 gctgccaccg ggacttgtcc acggtcagct tggagtacag gaagaaggag ccgtcggagt
     6121 ccagcacagg aggggtggtc ttgtagttgt tctcaggctg gccgttggac tcccactcca
     6181 cggcgatatc ggaagggtag aagcccttca ccagacaggt cagggacacc tggttcttgg
     6241 tcatttcctc cctgcttgga ggcagggtgt acacctgagg ctcgcgaggc tggcccttgg
     6301 ccttggagat ggtcttttcg atgggggcag gcagggcctt gttggagact ttgcacttgt
     6361 attccttgcc gttcagccag tcctggtgca gcacggtcag cacagacacc acccggtagg
     6421 tggagttgta ctgttcctcc cgaggcttgg tcttggcgtt gtgcacctc
//
