LOCUS       pEF.DEST51-PTPN1        7670 bp DNA     circular SYN 24-APR-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7670)
  AUTHORS   James
  TITLE     Direct Submission
  JOURNAL   Exported Mar 19, 2024 from SnapGene Viewer 7.1.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7670
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     promoter        475..1653
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          706..1644
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     promoter        1670..1688
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1720..1744
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1807..3577
                     /codon_start=1
                     /label=PTPN11
                     /note="Homo sapiens protein tyrosine phosphatase 
                     non-receptor type 11 (PTPN11), transcript variant 1, mRNA 
                     NCBI Reference Sequence: NM_002834.5"
                     /translation="MTSRRWFHPNITGVEAENLLLTRGVDGSFLARPSKSNPGDFTLSV
                     RRNGAVTHIKIQNTGDYYDLYGGEKFATLAELVQYYMEHHGQLKEKNGDVIELKYPLNC
                     ADPTSERWFHGHLSGKEAEKLLTEKGKHGSFLVRESQSHPGDFVLSVRTGDDKGESNDG
                     KSKVTHVMIRCQELKYDVGGGERFDSLTDLVEHYKKNPMVETLGTVLQLKQPLNTTRIN
                     AAEIESRVRELSKLAETTDKVKQGFWEEFETLQQQECKLLYSRKEGQRQENKNKNRYKN
                     ILPFDHTRVVLHDGDPNEPVSDYINANIIMPEFETKCNNSKPKKSYIATQGCLQNTVND
                     FWRMVFQENSRVIVMTTKEVERGKSKCVKYWPDEYALKEYGVMRVRNVKESAAHDYTLR
                     ELKLSKVGQGNTERTVWQYHFRTWPDHGVPSDPGGVLDFLEEVHHKQESIMDAGPVVVH
                     CSAGIGRTGTFIVIDILIDIIREKGVDCDIDVPKTIQMVRSQRSGMVQTEAQYRFIYMA
                     VQHYIETLQRRIEEEQKSKRKGHEYTNIKYSLADQTSGDQSPLPPCTPTPPCAEMREDS
                     ARVYENVGLMQQQK"
     protein_bind    complement(3606..3630)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3656..3697
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3707..3724
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3753..3977
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4023..4451
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4465..4795
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4646..4781
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4843..4890
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4909..5307
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5465..5586
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(6037..6625)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6796..7587,7588..7656))
                     /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"
     promoter        complement(join(7657..7670,1..91))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
       61 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
      121 tcgacggatc gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga
      181 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      241 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      301 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      361 ttgattattg actaggcttt tgcaaaaagc tttgcaaaga tggataaagt tttaaacaga
      421 gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgcctc gtgaggctcc
      481 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
      541 gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
      601 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
      661 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt
      721 ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc
      781 acctggctgc agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt
      841 tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcctggg
      901 cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat
      961 aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat
     1021 agtcttgtaa atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc
     1081 ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg
     1141 ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc
     1201 gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg
     1261 tgagcggaaa gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg
     1321 cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca
     1381 gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc
     1441 tcgagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt
     1501 ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc
     1561 ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt
     1621 caaagttttt ttcttccatt tcaggtgtcg tgaggaatta gcttggtact aatacgactc
     1681 actataggga gacccaagct ggctaggtaa gcttgatcaa caagtttgta caaaaaagca
     1741 ggctggcgcc ggaaccaatt cagtcgactg gatccggtac cgaggagatc tgccgccgcg
     1801 atcgccatga catcgcggag atggtttcac ccaaatatca ctggtgtgga ggcagaaaac
     1861 ctactgttga caagaggagt tgatggcagt tttttggcaa ggcctagtaa aagtaaccct
     1921 ggagacttca cactttccgt tagaagaaat ggagctgtca cccacatcaa gattcagaac
     1981 actggtgatt actatgacct gtatggaggg gagaaatttg ccactttggc tgagttggtc
     2041 cagtattaca tggaacatca cgggcaatta aaagagaaga atggagatgt cattgagctt
     2101 aaatatcctc tgaactgtgc agatcctacc tctgaaaggt ggtttcatgg acatctctct
     2161 gggaaagaag cagagaaatt attaactgaa aaaggaaaac atggtagttt tcttgtacga
     2221 gagagccaga gccaccctgg agattttgtt ctttctgtgc gcactggtga tgacaaaggg
     2281 gagagcaatg acggcaagtc taaagtgacc catgttatga ttcgctgtca ggaactgaaa
     2341 tacgacgttg gtggaggaga acggtttgat tctttgacag atcttgtgga acattataag
     2401 aagaatccta tggtggaaac attgggtaca gtactacaac tcaagcagcc ccttaacacg
     2461 actcgtataa atgctgctga aatagaaagc agagttcgag aactaagcaa attagctgag
     2521 accacagata aagtcaaaca aggcttttgg gaagaatttg agacactaca acaacaggag
     2581 tgcaaacttc tctacagccg aaaagagggt caaaggcaag aaaacaaaaa caaaaataga
     2641 tataaaaaca tcctgccctt tgatcatacc agggttgtcc tacacgatgg tgatcccaat
     2701 gagcctgttt cagattacat caatgcaaat atcatcatgc ctgaatttga aaccaagtgc
     2761 aacaattcaa agcccaaaaa gagttacatt gccacacaag gctgcctgca aaacacggtg
     2821 aatgactttt ggcggatggt gttccaagaa aactcccgag tgattgtcat gacaacgaaa
     2881 gaagtggaga gaggaaagag taaatgtgtc aaatactggc ctgatgagta tgctctaaaa
     2941 gaatatggcg tcatgcgtgt taggaacgtc aaagaaagcg ccgctcatga ctatacgcta
     3001 agagaactta aactttcaaa ggttggacaa gggaatacgg agagaacggt ctggcaatac
     3061 cactttcgga cctggccgga ccacggcgtg cccagcgacc ctgggggcgt gctggacttc
     3121 ctggaggagg tgcaccataa gcaggagagc atcatggatg cagggccggt cgtggtgcac
     3181 tgcagtgctg gaattggccg gacagggacg ttcattgtga ttgatattct tattgacatc
     3241 atcagagaga aaggtgttga ctgcgatatt gacgttccca aaaccatcca gatggtgcgg
     3301 tctcagaggt cagggatggt ccagacagaa gcacagtacc gatttatcta tatggcggtc
     3361 cagcattata ttgaaacact acagcgcagg attgaagaag agcagaaaag caagaggaaa
     3421 gggcacgaat atacaaatat taagtattct ctagcggacc agacgagtgg agatcagagc
     3481 cctctcccgc cttgtactcc aacgccaccc tgtgcagaaa tgagagaaga cagtgctaga
     3541 gtctatgaaa acgtgggcct gatgcaacag cagaaaagcg gccgctgata actcgagata
     3601 tctagaccca gctttcttgt acaaagtggt tgatctagag ggcccgcggt tcgaaggtaa
     3661 gcctatccct aaccctctcc tcggtctcga ttctacgcgt accggtcatc atcaccatca
     3721 ccattgagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     3781 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     3841 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     3901 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     3961 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     4021 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4081 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4141 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt
     4201 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4261 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4321 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4381 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4441 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4501 ccaggcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4561 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4621 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4681 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     4741 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     4801 cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg
     4861 gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagcct
     4921 ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc
     4981 atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact
     5041 ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact
     5101 gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg
     5161 ggcatcttga gcccctgcgg acggtgtcga caggtgcttc tcgatctgca tcctgggatc
     5221 aaagcgatag tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg
     5281 ccctctggtt atgtgtggga gggctaagca cttcgtggcc gaggagcagg actgacacgt
     5341 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5401 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5461 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     5521 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     5581 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     5641 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5701 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5761 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5821 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5881 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5941 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6001 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6061 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6121 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6181 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     6241 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6301 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6361 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6421 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     6481 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6541 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6601 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6661 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6721 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6781 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6841 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6901 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6961 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7021 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7081 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7141 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7201 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7261 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7321 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7381 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7441 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7501 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7561 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7621 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
//
