LOCUS       Exported                8554 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  To study localization and function of Tensin2.
ACCESSION   .
VERSION     .
KEYWORDS    pEGFP Tensin2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8554)
  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 8554)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8554
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      19..607
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     508..527
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        782..1085
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1086..1289
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1240..1260
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1328..1337
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1334..2050
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             1334..2050
                     /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(1379..1400)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1640..1659)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1987..2008
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    6063..6184
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6100..6119)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6154..6173
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6191..6646)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6328..6349)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6540..6559
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6673..6777
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6779..7136
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(6802..6822)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6987..7122
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7049..7068
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             7171..7965
                     /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(7225..7244)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7835..7854
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(8153..8172)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    8197..8244
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
       61 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
      121 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
      181 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      241 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      301 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      361 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      421 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      481 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      541 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      601 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      661 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
      721 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
      781 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      841 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      901 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      961 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     1021 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     1081 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     1141 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     1201 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     1261 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
     1321 accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1381 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     1441 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     1501 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     1561 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     1621 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     1681 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     1741 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     1801 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     1861 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1921 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     1981 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     2041 gctgtacaag tccggactca gatctcgagc tcaagcttcg aattctatgg agcggcgctg
     2101 ggacttagac ctcacctacg tgacggagcg catcttggcc gccgccttcc ccgcgcggcc
     2161 cgatgaacag cggcaccggg gccacctgcg cgagctggcc catgtgctgc aatccaagca
     2221 ccgggacaag tacctgctct tcaacctttc agagaaaagg catgacctga cccgcttaaa
     2281 ccccaaggtt caagacttcg gctggcctga gctgcatgct ccacccctgg acaagctgtg
     2341 ctccatctgc aaagccatgg agacatggct cagtgctgac ccacagcacg tggtcgtact
     2401 atactgcaag ggaaacaagg gcaagcttgg ggtcatcgtt tctgcctaca tgcactacag
     2461 caagatctct gcaggggcgg accaggcact ggccactctt accatgcgga aattctgcga
     2521 ggacaaggtg gccacagaac tgcagccctc ccagcgtcga tatatcagct acttcagtgg
     2581 gctgctatct ggctccatca gaatgaacag cagccctctc ttcctgcact atgtgctcat
     2641 ccccatgctg ccagcctttg aacctggcac aggcttccag cccttcctta aaatctacca
     2701 gtccatgcag cttgtctaca catctggagt ctatcacatt gcaggccctg gtccccagca
     2761 gctttgcatc agcctggagc cagccctcct cctcaaaggc gatgtcatgg taacatgtta
     2821 tcacaagggt ggccggggca cagaccggac cctcgtgttc cgagtccagt tccacacctg
     2881 caccatccac ggaccacagc tcactttccc caaggaccag cttgacgagg cctggactga
     2941 tgagaggttc cccttccaag cctccgtgga gtttgtcttc tcctccagcc ccgagaagat
     3001 caaaggcagc actccacgga acgacccctc ggtctctgtc gactacaaca ccactgagcc
     3061 agccgtgcgc tgggactcct atgagaactt caaccagcac cacgaggaca gtgtggatgg
     3121 ctccttgacc cacacccggg gtcccctgga tggcagtcct tatgcccagg tgcagcggcc
     3181 tccccggcag acccccccgg caccctctcc agagcctcca ccacccccca tgctctctgt
     3241 cagcagcgac tcaggccatt cctccacgct gaccacagag ccggctgctg agtcccctgg
     3301 ccggccgccc cctacagctg ctgaacggca ggagctggat cgcctcctag gaggctgcgg
     3361 agtggccagt gggggccggg gagctgggcg cgagacggcc atcctagatg acgaagagca
     3421 gcccactgtg ggcggaggcc cccacctcgg agtgtatcca ggccataggc ctggcctcag
     3481 ccgccactgc tcctgccgcc agggctaccg ggagccctgc ggggttccca atgggggcta
     3541 ctaccggcca gagggaaccc tggagaggag gcgactggcc tacgggggct atgagggatc
     3601 cccccagggc tacgccgagg cctcgatgga gaagaggcgc ctctgccgat cgctgtcaga
     3661 ggggctatac ccctacccac ctgagatggg gaaaccagcc actggggact ttggctaccg
     3721 cgccccaggc taccgggagg tggtcatcct ggaggaccct gggctgcctg ccctataccc
     3781 atgcccagcc tgcgaggaga agctggcgct gcctacagca gccttgtatg gactgcggct
     3841 ggagagggag gctggagaag ggtgggcaag tgaggctggc aagcctctcc tgcacccagt
     3901 gcggcctggg cacccgctgc ctctgctctt gcctgcctgt gggcatcacc atgccccgat
     3961 gcctgactac agctgcctga agccacccaa ggcaggcgag gaagggcacg agggctgctc
     4021 ctacaccatg tgccccgaag gcaggtatgg gcatccaggg taccctgccc tggtgacata
     4081 cagctatgga ggagcagttc ccagttactg cccagcatat ggccgtgtgc ctcatagctg
     4141 tggctctcca ggagagggca gagggtatcc cagccctggt gcccactccc cacgggctgg
     4201 ctccatttcc ccgggcagcc cgccctatcc acaatctagg aagctgagct acgagatccc
     4261 tacggaggag ggaggggaca ggtacccatt gcctgggcac ctggcctcag caggaccttt
     4321 ggcatctgca gagtcgctgg agccggtgtc ctggagggag ggccccagtg ggcacagcac
     4381 actgcctcgg tctccccgag atgccccatg cagtgcttcg tcagagttgt ctggtccctc
     4441 cacgcccctg cacaccagca gtccagtcca gggcaaggaa agcacccggc gacaggacac
     4501 caggtccccc acctcagcgc ccactcagag actgagtcct ggcgaggcct tgccccctgt
     4561 ttcccaggca ggcaccggaa aggcccctga gctgccgtcg ggaagtgggc ctgagcctct
     4621 ggcccctagc ccagtctctc cgaccttccc tcccagctcg cccagtgact ggcctcagga
     4681 aaggagtcca gggggccact cagatggcgc cagtcctcgg agccctgtgc ccaccacact
     4741 tcctggcctc cgccacgccc cctggcaagg ccctcgaggc ccccccgaca gcccagatgg
     4801 gtctcccctc actcctgtgc cttcccagat gccctggctt gtggccagcc cagagccgcc
     4861 tcagagctca cctacacctg ctttccccct ggctgcctcc tatgacacca atggccttag
     4921 ccagccccca cttcctgaga aacgccacct gcccgggccg gggcaacagc caggaccctg
     4981 gggcccagag caggcatcat cgccagccag aggcatcagt caccatgtca ccttcgcacc
     5041 tctgctctca gataatgtcc cccaaacccc agagcctcct acacaagaga gccaaagcaa
     5101 tgtcaagttt gtccaggata catccaagtt ctggtacaag ccacacctgt cccgtgacca
     5161 agccattgcc ctgctgaagg acaaggaccc tggggccttc ctgatcaggg acagtcattc
     5221 attccaagga gcttatgggc tggccctcaa ggtggccaca ccgccaccca gtgcccagcc
     5281 ctggaaaggg gaccccgtgg aacagctggt ccgccatttc ctcatcgaga ctgggcccaa
     5341 aggggtgaag atcaagggct gccccagtga gccctacttt ggcagcctgt ccgccttggt
     5401 ctcccagcac tccatctccc ccatctccct gccctgctgc ctgcgcattc ccagcaaaga
     5461 tcctctggaa gagaccccag aggctccagt gcccaccaac atgagcacag cggcagacct
     5521 cctgcgtcag ggtgctgcct gcagcgtgct ctacttgacc tcagtggaga cagagtcact
     5581 gacgggcccc caagctgtgg cccgggccag ctctgcagct ctgagctgta gcccccgccc
     5641 gacaccagct gttgtccact tcaaggtgtc agcccagggc attacactga cggacaacca
     5701 aaggaagctc ttctttcgcc gccattatcc agtgaacagc atcaccttct ccagcactga
     5761 ccctcaagac cggagatgga ccaacccaga cgggaccacc tccaagatct ttggtttcgt
     5821 ggccaagaag ccgggaagcc cctgggagaa tgtgtgtcac ctctttgcag agcttgaccc
     5881 agatcagcct gctggcgcca ttgtcacctt catcaccaaa gttctactgg gccagagaaa
     5941 atgaatctag ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     6001 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     6061 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     6121 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     6181 cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     6241 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     6301 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     6361 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     6421 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     6481 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     6541 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     6601 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     6661 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     6721 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     6781 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     6841 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     6901 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     6961 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     7021 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     7081 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     7141 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     7201 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     7261 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     7321 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     7381 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     7441 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     7501 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     7561 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     7621 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     7681 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     7741 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     7801 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     7861 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     7921 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     7981 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     8041 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     8101 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     8161 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     8221 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     8281 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     8341 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     8401 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     8461 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     8521 agttttcgtt ccactgagcg tcagaccccg taga
//
