LOCUS       Exported                4595 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Fluid Shear Stress inducible Red Fluorescent Protein for mammalian 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    EGR-1 TurboRFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4595)
  AUTHORS   Varma S, Voldman J
  TITLE     A cell-based sensor of fluid shear stress for microfluidics.
  JOURNAL   Lab Chip. 2015 Feb 4.
  PUBMED    25648195
REFERENCE   2  (bases 1 to 4595)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4595
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             984..995
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             1268..1963
                     /codon_start=1
                     /product="red fluorescent protein from Entacmaea 
                     quadricolor"
                     /label=TurboRFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     polyA_signal    2086..2207
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2123..2142)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2177..2196
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2214..2669)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2351..2372)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2563..2582
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2696..2800
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2802..3159
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2825..2845)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3010..3145
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3072..3091
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3194..3988
                     /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(3248..3267)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3858..3877
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4176..4195)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4220..4267
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat tactagcgct
      721 accggactca gatctcgagc tggccctccc cacgcgggcg tccccgactc ccgcgcgcgc
      781 tcaggctccc agttgggaac caaggagggg gaggatgggg gggggggtgt gcgccgaccc
      841 ggaaacgcca tataaggagc aggaaggatc ccccgccgga acagacctta tttgggcagc
      901 gccttatatg gagtggccca atatggccct gccgcttccg gctctgggag gaggggcgag
      961 cgggggttgg ggcgggggca agctgggaac tccaggcgcc tggcccggga ggccactgct
     1021 gctgttccaa tactaggctt tccaggagcc tgagcgctcg cgatgccgga gcgggtcgca
     1081 gggtggaggt gcccaccact cttggatggg agggcttcac gtcactccgg gtcctcccgg
     1141 ccggtccttc catattaggg cttcctgctt cccatatatg gccatgtacg tcacggcgga
     1201 ggcgggcccg tgctgttcca gacccttgaa atagaggccg attcggggag tcgcaccggt
     1261 cgccaccatg agcgagctga tcaaggagaa catgcacatg aagctgtaca tggagggcac
     1321 cgtgaacaac caccacttca agtgcacatc cgagggcgaa ggcaagccct acgagggcac
     1381 ccagaccatg aagatcaagg tggtcgaggg cggccctctc cccttcgcct tcgacatcct
     1441 ggctaccagc ttcatgtacg gcagcaaagc cttcatcaac cacacccagg gcatccccga
     1501 cttctttaag cagtccttcc ctgagggctt cacatgggag agaatcacca catacgaaga
     1561 cgggggcgtg ctgaccgcta cccaggacac cagcttccag aacggctgca tcatctacaa
     1621 cgtcaagatc aacggggtga acttcccatc caacggccct gtgatgcaga agaaaacacg
     1681 cggctgggag gccaacaccg agatgctgta ccccgctgac ggcggcctga gaggccacag
     1741 ccagatggcc ctgaagctcg tgggcggggg ctacctgcac tgctccttca agaccacata
     1801 cagatccaag aaacccgcta agaacctcaa gatgcccggc ttccacttcg tggaccacag
     1861 actggaaaga atcaaggagg ccgacaaaga gacctacgtc gagcagcacg agatggctgt
     1921 ggccaagtac tgcgacctcc ctagcaaact ggggcacaga tgatgcggcc gcgactctag
     1981 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     2041 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     2101 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     2161 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     2221 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     2281 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     2341 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     2401 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     2461 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     2521 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     2581 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     2641 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     2701 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2761 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     2821 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2881 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2941 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3001 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3061 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     3121 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     3181 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3241 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     3301 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     3361 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     3421 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     3481 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     3541 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     3601 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     3661 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     3721 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     3781 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     3841 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     3901 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     3961 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     4021 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4081 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4141 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     4201 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     4261 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     4321 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     4381 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     4441 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4501 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4561 gcgtcagacc ccgtagaaaa gatcaaagga tcttc
//
