LOCUS       Exported                6361 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  In vivo visualization of focal adhesions (can be used for 
            colocalization studies).
ACCESSION   .
VERSION     .
KEYWORDS    mScarlet-I-PXN
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6361)
  AUTHORS   Chertkova AO, Mastop M, Postma M, van Bommel N, van der Niet S, 
            Batenburg KL, Joosen L, Gadella TWJ, Okada Y, Goedhart J
  TITLE     Robust and Bright Genetically Encoded Fluorescent Markers for 
            Highlighting Structures and Compartments in Mammalian Cells
  JOURNAL   bioRxiv 160374
REFERENCE   2  (bases 1 to 6361)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6361
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             613..1308
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(746..764)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1014..1033
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3149..3270
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3186..3205)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3240..3259
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3277..3732)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3414..3435)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3626..3645
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3759..3863
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3865..4222
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3888..3908)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4073..4208
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4135..4154
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4257..5051
                     /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="MIEQDGLHEGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTESGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4311..4330)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4921..4940
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5239..5258)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5283..5330
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5659..6247
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6148..6167
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gcagtgatca aggagttcat gcggttcaag
      661 gtgcacatgg agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc
      721 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
      781 ttctcctggg acatcctgtc ccctcagttc atgtacggct ccagggcctt catcaagcac
      841 cccgccgaca tccccgacta ctataagcag tccttccccg agggcttcaa gtgggagcgc
      901 gtgatgaact tcgaggacgg cggcgccgtg accgtgaccc aggacacctc cctggaggac
      961 ggcaccctga tctacaaggt gaagctccgc ggcaccaact tccctcctga cggccccgta
     1021 atgcagaaga agacaatggg ctgggaagcg tccaccgagc ggttgtaccc cgaggacggc
     1081 gtgctgaagg gcgacattaa gatggccctg cgcctgaagg acggcggccg ctacctggcg
     1141 gacttcaaga ccacctacaa ggccaagaag cccgtgcaga tgcccggcgc ctacaacgtc
     1201 gaccgcaagt tggacatcac ctcccacaac gaggactaca ccgtggtgga acagtacgaa
     1261 cgctccgagg gccgccactc caccggcggc atggacgagc tgtacaagtc cggactcaga
     1321 tctcgagctg cggccgccgg atccaagctt atggacgacc tcgacgccct gctggcggac
     1381 ttggagtcta ccacctccca catctccaaa cggcctgtgt tcttgtcgga ggagaccccc
     1441 tactcatacc caactggaaa ccacacatac caggagattg ccgtgccacc ccccgtcccc
     1501 ccacccccgt ccagcgaggc cctcaatggc acaatccttg accccttaga ccagtggcag
     1561 cccagcggct cccgattcat ccaccagcag cctcagtcct catcacctgt gtacggctcc
     1621 agtgccaaaa cttccagtgt ctccaaccct caggacagtg ttggctctcc gtgctcccga
     1681 gtgggtgagg aggagcacgt ctacagcttc cccaacaagc agaaatcagc tgagccttca
     1741 cccaccgtaa tgagcacgtc cctgggcagc aacctttctg aactcgaccg cctgctgctg
     1801 gaactgaacg ctgtacagca taacccgcca ggcttccctg cagatgaggc caactcaagc
     1861 cccccgcttc ctggggccct gagccccctc tatggtgtcc cagagactaa cagccccttg
     1921 ggaggcaaag ctgggcccct gacgaaagag aagcctaagc ggaatggggg ccggggcctg
     1981 gaggacgtgc ggcccagtgt ggagagtctc ttggatgaac tggagagctc cgtgcccagc
     2041 cccgtccctg ccatcactgt gaaccagggc gagatgagca gcccgcagcg cgtcacctcc
     2101 acccaacagc agacacgcat ctcggcctcc tctgccacca gggagctgga cgagctgatg
     2161 gcttcgctgt cggatttcaa gttcatggcc caggggaaga cagggagcag ctcaccccct
     2221 ggggggcccc cgaagcccgg gagccagctg gacagcatgc tggggagcct gcagtctgac
     2281 ctgaacaagc tgggggtcgc cacagtcgcc aaaggagtct gcggggcctg caagaagccc
     2341 atcgccgggc aggttgtgac cgccatgggg aagacgtggc accccgagca cttcgtctgc
     2401 acccactgcc aggaggagat cggatcccgg aacttcttcg agcgggatgg acagccctac
     2461 tgtgaaaagg actaccacaa cctcttctcc ccgcgctgct actactgcaa cggccccatc
     2521 ctggataaag tggtgacagc ccttgaccgg acgtggcacc ctgaacactt cttctgtgca
     2581 cagtgtggag ccttctttgg tcccgaaggg ttccacgaga aggacggcaa ggcctactgt
     2641 cgcaaggact acttcgacat gttcgcaccc aagtgtggcg gctgcgcccg ggccatcctg
     2701 gagaactata tctcagccct caacacgctg tggcatcctg agtgctttgt gtgccgggaa
     2761 tgcttcacgc cattcgtgaa cggcagcttc ttcgagcacg acgggcagcc ctactgtgag
     2821 gtgcactacc acgagcggcg cggctcgctg tgttctggct gccagaagcc catcaccggc
     2881 cgctgcatca ccgccatggc caagaagttc caccccgagc acttcgtctg tgccttctgc
     2941 ctcaagcagc tcaacaaggg caccttcaag gagcagaacg acaagcctta ctgtcagaac
     3001 tgcttcctca agctcttctg ctaggctcga gtctagataa ctgatcataa tcagccatac
     3061 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     3121 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     3181 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3241 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     3301 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     3361 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     3421 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     3481 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     3541 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     3601 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     3661 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3721 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3781 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3841 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     3901 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3961 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4021 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4081 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4141 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     4201 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     4261 ttgaacaaga tggattgcac gaaggttctc cggccgcttg ggtggagagg ctattcggct
     4321 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4381 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     4441 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4501 acgttgtcac tgaatcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4561 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4621 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4681 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4741 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     4801 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4861 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4921 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4981 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     5041 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     5101 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5161 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5221 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     5281 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     5341 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     5401 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     5461 cgcagccaac gtcggggcgg caggcactgc catagcctca ggttactcat atatacttta
     5521 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5581 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5641 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5701 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5761 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5821 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5881 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5941 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6001 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6061 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6121 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6181 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6241 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6301 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     6361 t
//
