LOCUS       Exported                9055 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  To study localization and function of Tensin3.
ACCESSION   .
VERSION     .
KEYWORDS    pEGFP Tensin3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9055)
  AUTHORS   Clark K, Howe JD, Pullar CE, Green JA, Artym VV, Yamada KM, 
            Critchley DR
  TITLE     Tensin 2 modulates cell contractility in 3D collagen gels through 
            the RhoGAP DLC1.
  JOURNAL   J Cell Biochem. 2010 Mar 1;109(4):808-17. doi: 10.1002/jcb.22460.
  PUBMED    20069572
REFERENCE   2  (bases 1 to 9055)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9055
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      6..594
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     495..514
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        758..1061
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1062..1265
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1216..1236
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1304..1313
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1310..2026
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             1310..2026
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1355..1376)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1616..1635)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1963..1984
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    6551..6672
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6588..6607)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6642..6661
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6679..7134)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6816..6837)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7028..7047
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        7161..7265
                     /gene="bla"
                     /label=AmpR promoter
     promoter        7267..7624
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(7290..7310)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      7475..7610
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7537..7556
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             7659..8453
                     /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(7713..7732)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8323..8342
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(8641..8660)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    8685..8732
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
       61 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
      121 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
      181 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
      241 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
      301 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
      361 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
      421 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
      481 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
      541 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
      601 agcaacgcgg cctttttacg gttcctggcc ttttgctcac atgttctttc ctgcgttatc
      661 ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta
      721 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      781 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      841 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      901 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      961 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     1021 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     1081 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     1141 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     1201 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     1261 gagctggttt agtgaaccgt cagatccgct agcgctaccg gtcgccacca tggtgagcaa
     1321 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     1381 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     1441 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1501 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     1561 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     1621 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1681 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     1741 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     1801 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     1861 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     1921 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1981 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtccg gactcagatc
     2041 tcgagctcaa gcttcgaatt ccatggagga gggccatggg ctggacctca cttacatcac
     2101 ggagcgcatc atcgctgtgt ccttccctgc cggctgctct gaggagtcct acctgcacaa
     2161 cctacaggag gtcacgcgca tgctcaagtc caagcacggg gacaactacc tggtattaaa
     2221 cctttcagaa aagagatatg accttacgaa gcttaaccca aagatcatgg atgtgggctg
     2281 gccagagctc cacgcaccgc ccctggataa gatgtgtacc atatgcaagg cgcaggagtc
     2341 ctggctgaac agcaacctcc agcatgtggt cgtcattcac tgcaggggcg ggaaaggacg
     2401 cataggagtg gtcatatcat cctacatgca tttcaccaac gtctcagcca gcgccgacca
     2461 ggcccttgac aggtttgcaa tgaagaagtt ttatgatgac aaagtttcag ctttaatgca
     2521 gccttcccaa aaacggtatg ttcagttcct cagtgggctc ctgtccggat cggtgaaaat
     2581 gaatgcctct cccctgttcc tgcattttgt catcctccac ggcaccccca acttcgacac
     2641 aggtggagtg tgccggccct ttctgaagct ctaccaagcc atgcagcctg tgtacacctc
     2701 cgggatctac aacgttggcc cagaaaaccc cagcaggatc tgcatcgtca tcgagccggc
     2761 ccagcttctg aagggagatg tcatggtgaa atgctaccac aagaaatacc gctcggccac
     2821 ccgtgacgtc attttccgcc tgcagtttca cactggggct gtgcagggct acgggctggt
     2881 gtttgggaag gaggatctgg acaatgccag caaagatgac cgttttcctg actatgggaa
     2941 ggttgaatta gtcttctctg ccacgcctga gaagattcaa gggtccgaac acttgtacaa
     3001 cgaccacggt gtgattgtgg actacaacac aacagaccca ctgatacgct gggactcgta
     3061 cgagaacctc agtgcagatg gagaagtgct acacacgcag ggccctgtcg atggcagcct
     3121 ttacgcgaag gtgaggaaga aaagctcctc ggatcctggc atcccaggtg gcccccaggc
     3181 aatcccggcc accaacagcc cagaccacag tgaccacacc ttgtctgtca gcagtgactc
     3241 cggccactct acagcctctg ccaggacgga taagacggaa gagcgcctgg ccccaggaac
     3301 caggaggggc ctgagtgccc aggagaaggc tgagttggac cagctgctca gtggctttgg
     3361 cctggaagat cctggaagct ccctcaagga aatgactgat gctcgaagca agtacagtgg
     3421 gacccgccac gtggtgccag cccaggttca cgtgaatgga gacgctgctc tgaaggatcg
     3481 ggagacagac attctggatg acgagatgcc ccaccacgac ctgcacagtg tggacagcct
     3541 tgggaccctg tcctcctcgg aagggcctca gtcggcccac ctgggtccct tcacctgcca
     3601 caagagcagc cagaactcac tcctatctga cggttttggc agcaacgttg gtgaagatcc
     3661 gcagggcacc ctcgttccgg acctgggcct tggcatggac ggcccctatg agcgggagcg
     3721 gacttttggg agtcgagagc ccaagcagcc ccagcccctg ctgagaaagc cctcagtgtc
     3781 cgcccagatg caggcctatg ggcagagcag ctactccaca cagacctggg tgcgccagca
     3841 gcagatggtt gtagctcacc agtatagctt cgccccagat ggggaggccc ggctggtgag
     3901 ccgctgccct gcagacaatc ctggcctcgt ccaggcccag cccagagtgc cactcacccc
     3961 cacccgaggg accagcagta gggtggctgt ccagaggggt gtaggcagtg ggccacatcc
     4021 ccctgacaca cagcagccct ctcccagcaa agcgttcaaa cccaggtttc caggagacca
     4081 ggttgtgaat ggagccggcc cagagctgag cacaggcccc tccccaggct cgcccaccct
     4141 ggacatcgac cagtccatcg agcagctcaa caggctgatc ctggagctgg atcccacctt
     4201 cgagcccatc cctacccaca tgaacgccct cggtagccag gccaatggct ctgtgtctcc
     4261 agacagcgtg ggaggcgggc tccgggcaag cagcaggctg cctgacacag gagagggccc
     4321 cagcagggcc accgggcggc aaggctcctc tgctgaacag cccctgggcg ggagactcag
     4381 gaagctgagc ctggggcagt acgacaacga tgctgggggg cagctgccct tctccaaatg
     4441 tgcatgggga aaggctggtg tggactatgc cccaaacctg ccgccattcc cctcaccagc
     4501 ggacgtcaaa gagacgatga cccctggcta tccccaggac ctcgatatta tcgatggcag
     4561 aattttaagt agcaaggagt ccatgtgttc aactccagca tttcctgtgt ctccagagac
     4621 accgtatgtg aaaacagcgc tgcgccatcc tccgttcagc ccacctgagc ccccgctgag
     4681 cagcccagcc agtcagcaca aaggaggacg tgaaccacga agctgccctg agacgctcac
     4741 tcacgctgtg gggatgtcag agagccccat cggacccaaa tccacgatgc tccgggctga
     4801 tgcgtcctcg acgccctcct ttcagcaggc ttttgcttct tcctgcacca tttccagcaa
     4861 cggccctggg cagaggagag agagctcctc ttctgcagaa cgccagtggg tggagagcag
     4921 ccccaagccc atggtttccc tgctggggag cggccggccc accggaagtc ccctcagcgc
     4981 tgagttctcc ggtaccagga aggactcccc agtgctgtcc tgcttcccgc cgtcagagct
     5041 ccaggctcct ttccacagcc atgagctgtc cctagcagag ccaccggact ccctggcgcc
     5101 tcccagcagc caggccttcc tgggcttcgg caccgcccca gtgggaagtg gccttccgcc
     5161 cgaggaggac ctgggggcct tgctggccaa ttctcatgga gcgtcaccga cccccagcat
     5221 cccgctgaca gcgacagggg ctgccgacaa tggcttcctg tcccacaact ttctcacggt
     5281 ggcgcctgga cacagcagcc accacagtcc aggcctgcag ggccagggtg tgaccctgcc
     5341 cgggcagcca cccctccctg agaagaagcg ggcctcggag ggggatcgtt ctttgggctc
     5401 agtctctccc tcctccagtg gcttctccag cccgcacagc gggagcacca tcagtatccc
     5461 cttcccaaat gtccttcccg acttttccaa ggcttcagaa gcggcctcac ctctgccaga
     5521 tagtccaggt gataaacttg tgatcgtgaa atttgttcaa gacacttcca agttctggta
     5581 caaggcggat atttcaagag aacaagccat cgccatgttg aaggacaagg agccgggctc
     5641 attcattgtt cgagacagcc attccttccg aggggcctat ggcctggcca tgaaggtggc
     5701 cacgccccca ccttcagtcc tgcagctgaa caagaaagct ggagatttgg ccaatgaact
     5761 cgtccggcac tttttgatcg agtgtacccc gaagggagtg cggttgaaag ggtgctcgaa
     5821 tgaaccatat ttcgggagcc tgacggcctt ggtgtgccag cattccatca cgcccttggc
     5881 cttgccgtac aagctgctta tcccagagag agatccattg gaggaaatag cagaaagttc
     5941 tccccagacg gcagccaatt cagcagctga gctgttgaag cagggggcag cctgcaatgt
     6001 gtggtacttg aactctgtgg agatggagtc cctcaccggc caccaggcga tccagaaggc
     6061 cctgagcatc accctggtcc aggagcctcc acctgtgtcc acagttgtgc acttcaaggt
     6121 gtcagcccag ggcatcaccc tgacagacaa tcagaggaag ctcttcttcc ggaggcatta
     6181 ccccgtgaac agtgtgattt tctgtgcctt ggacccacaa gacaggaagt ggatcaaaga
     6241 tggcccttcc tcaaaagtct ttggatttgt ggcccggaag cagggcagtg ccacggataa
     6301 tgtgtgccac ctgtttgcag agcatgaccc tgagcagcct gccagtgcca ttgtcaactt
     6361 cgtatcaaag gtcatgattg gttcaccaaa gaaggtctga gtcgacggta ccgcgggccc
     6421 gggatccacc ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt
     6481 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     6541 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     6601 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     6661 caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     6721 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     6781 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     6841 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     6901 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     6961 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     7021 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     7081 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     7141 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     7201 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     7261 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     7321 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     7381 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     7441 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     7501 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     7561 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     7621 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     7681 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     7741 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     7801 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     7861 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     7921 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     7981 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     8041 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     8101 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     8161 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     8221 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     8281 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     8341 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     8401 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     8461 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     8521 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     8581 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     8641 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     8701 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     8761 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     8821 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     8881 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     8941 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     9001 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aagga
//
