LOCUS       Exported                6024 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  In vivo visualization of microtubules (can be used for 
            colocalization studies).
ACCESSION   .
VERSION     .
KEYWORDS    pmScarlet_alphaTubulin_C1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6024)
  AUTHORS   Bindels DS, Haarbosch L, van Weeren L, Postma M, Wiese KE, Mastop M,
            Aumonier S, Gotthard G, Royant A, Hink MA, Gadella TW Jr
  TITLE     mScarlet: a bright monomeric red fluorescent protein for cellular 
            imaging.
  JOURNAL   Nat Methods. 2016 Nov 21. doi: 10.1038/nmeth.4074.
  PUBMED    27869816
REFERENCE   2  (bases 1 to 6024)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6024
                     /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
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     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    2812..2933
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2849..2868)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2903..2922
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2940..3395)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3077..3098)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3289..3308
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3422..3526
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3528..3885
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3551..3571)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3736..3871
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3798..3817
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3920..4714
                     /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(3974..3993)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4584..4603
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4902..4921)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4946..4993
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5322..5910
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5811..5830
                     /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 caccaagcac
      841 cccgccgaca tccccgacta ctataagcag tccttccccg agggcttcaa gtgggagcgc
      901 gtgatgaact tcgaggacgg cggcgccgtg accgtgaccc aggacacctc cctggaggac
      961 ggcaccctga tctacaaggt gaagctccgc ggcaccaact tccctcctga cggccccgta
     1021 atgcagaaga agacaatggg ctgggaagca 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 tctcgagtgc gtgagtgcat ctccatccac gttggccagg ctggtgtcca gattggcaat
     1381 gcctgctggg agctctactg cctggaacac ggcatccagc ccgatggcca gatgccaagt
     1441 gacaagacca ttgggggagg agatgactcc ttcaacacct tcttcagtga gacgggcgct
     1501 ggcaagcacg tgccccgggc tgtgtttgta gacttggaac ccacagtcat tgatgaagtt
     1561 cgcactggca cctaccgcca gctcttccac cctgagcagc tcatcacagg caaggaagat
     1621 gctgccaata actatgcccg agggcactac accattggca aggagatcat tgaccttgtg
     1681 ttggaccgaa ttcgcaagct ggctgaccag tgcaccggtc ttcagggctt cttggttttc
     1741 cacagctttg gtgggggaac tggttctggg ttcacctccc tgctcatgga acgtctctca
     1801 gttgattatg gcaagaagtc caagctggag ttctccattt acccagcacc ccaggtttcc
     1861 acagctgtag ttgagcccta caactccatc ctcaccaccc acaccaccct ggagcactct
     1921 gattgtgcct tcatggtaga caatgaggcc atctatgaca tctgtcgtag aaacctcgat
     1981 atcgagcgcc caacctacac taaccttaac cgccttatta gccagattgt gtcctccatc
     2041 actgcttccc tgagatttga tggagccctg aatgttgacc tgacagaatt ccagaccaac
     2101 ctggtgccct acccccgcat ccacttccct ctggccacat atgcccctgt catctctgct
     2161 gagaaagcct accatgaaca gctttctgta gcagagatca ccaatgcttg ctttgagcca
     2221 gccaaccaga tggtgaaatg tgaccctcgc catggtaaat acatggcttg ctgcctgttg
     2281 taccgtggtg acgtggttcc caaagatgtc aatgctgcca ttgccaccat caaaaccaag
     2341 cgcagcatcc agtttgtgga ttggtgcccc actggcttca aggttggcat caactaccag
     2401 cctcccactg tggtgcctgg tggagacctg gccaaggtac agagagctgt gtgcatgctg
     2461 agcaacacca cagccattgc tgaggcctgg gctcgcctgg accacaagtt tgacctgatg
     2521 tatgccaagc gtgcctttgt tcactggtac gtgggtgagg ggatggagga aggcgagttt
     2581 tcagaggccc gtgaagatat ggctgccctt gagaaggatt atgaggaggt tggtgtggat
     2641 tctgttgaag gagagggtga ggaagaagga gaggaatact aaggatccac cggatctaga
     2701 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     2761 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     2821 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2881 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta
     2941 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     3001 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     3061 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     3121 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     3181 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     3241 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     3301 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     3361 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     3421 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3481 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     3541 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3601 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3661 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3721 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3781 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3841 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     3901 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3961 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4021 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4081 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4141 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4201 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4261 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4321 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4381 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4441 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4501 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4561 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4621 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4681 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4741 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4801 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4861 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     4921 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     4981 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     5041 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     5101 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     5161 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5221 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5281 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5341 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5401 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5461 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5521 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5581 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5641 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5701 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5761 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5821 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5881 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5941 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6001 gataaccgta ttaccgccat gcat
//
