LOCUS       sequence-310007-        5440 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  membrane marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5440)
  AUTHORS   Bisaria A, Hayer A, Garbett D, Cohen D, Meyer T
  TITLE     Membrane-proximal F-actin restricts local membrane protrusions and 
            directs cell migration.
  JOURNAL   Science. 2020 Jun 12;368(6496):1205-1210. doi: 
            10.1126/science.aay7794.
  PUBMED    32527825
REFERENCE   2  (bases 1 to 5440)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2020-06-12- 12"; volume: "368"; 
            issue: "6496"; pages: "1205-1210"
COMMENT     Sequence Label: C1-iRFP-iLid-CaaX
FEATURES             Location/Qualifiers
     source          1..5440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        64..367
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        368..571
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     522..542
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             631..1578
                     /codon_start=1
                     /product="phytochrome-based near-infrared fluorescent 
                     protein, originally termed iRFP (Filonov et al., 2011)"
                     /label=iRFP713
                     /note="name was changed to iRFP713 (Shcherbakova and 
                     Verkhusha, 2013)"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     CDS             1609..2046
                     /codon_start=1
                     /product="improved light-inducible dimer when used with 
                     SspB nano or SspB micro (Guntas et al., 2015)"
                     /label=iLID
                     /translation="LATTLERIEKNFVITDPRLPDNPIIFASDSFLQLTEYSREEILGR
                     NCRFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNVFHLQPMRDYKGDVQ
                     YFIGVQLDGTERLHGAAEREAVCLIKKTAFQIAEAANDENYF"
     polyA_signal    2231..2352
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2268..2287)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2322..2341
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2359..2814)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2496..2517)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2708..2727
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2841..2945
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2947..3304
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2947..3138
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2970..2990)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3155..3290
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3339..4133
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3339..4133
                     /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(3393..3412)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4003..4022
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4321..4340)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4365..4412
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4741..5329
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5230..5249
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
       61 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      121 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      181 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      241 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      301 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      361 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      421 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      481 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      541 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg
      601 ctaccggact cagatctcga gctcaagctt atggctgaag gatccgtcgc caggcagcct
      661 gacctcttga cctgcgacga tgagccgatc catatccccg gtgccatcca accgcatgga
      721 ctgctgctcg ccctcgccgc cgacatgacg atcgttgccg gcagcgacaa ccttcccgaa
      781 ctcaccggac tggcgatcgg cgccctgatc ggccgctctg cggccgatgt cttcgactcg
      841 gagacgcaca accgtctgac gatcgccttg gccgagcccg gggcggccgt cggagcaccg
      901 atcactgtcg gcttcacgat gcgaaaggac gcaggcttca tcggctcctg gcatcgccat
      961 gatcagctca tcttcctcga gctcgagcct ccccagcggg acgtcgccga gccgcaggcg
     1021 ttcttccgcc gcaccaacag cgccatccgc cgcctgcagg ccgccgaaac cttggaaagc
     1081 gcctgcgccg ccgcggcgca agaggtgcgg aagattaccg gcttcgatcg ggtgatgatc
     1141 tatcgcttcg cctccgactt cagcggcgaa gtgatcgcag aggatcggtg cgccgaggtc
     1201 gagtcaaaac taggcctgca ctatcctgcc tcaaccgtgc cggcgcaggc ccgtcggctc
     1261 tataccatca acccggtacg gatcattccc gatatcaatt atcggccggt gccggtcacc
     1321 ccagacctca atccggtcac cgggcggccg attgatctta gcttcgccat cctgcgcagc
     1381 gtctcgcccg tccatctgga attcatgcgc aacataggca tgcacggcac gatgtcgatc
     1441 tcgattttgc gcggcgagcg actgtgggga ttgatcgttt gccatcaccg aacgccgtac
     1501 tacgtcgatc tcgatggccg ccaagcctgc gagctagtcg cccaggttct ggcctggcag
     1561 atcggcgtga tggaagagct gtacaagggt agtggtagtg gggagtttct ggcaaccaca
     1621 ctggaacgga tcgagaaaaa tttcgtgatt actgatccga gactgcctga caacccaatc
     1681 atttttgcga gcgattcctt cctgcagctg acagaatatt ctcgggaaga gatcctgggg
     1741 cgcaattgcc gttttctgca gggacccgag acagaccgtg ccactgttcg gaaaatcaga
     1801 gatgctattg acaaccagac tgaagtgacc gttcagctga tcaattatac caagagcggc
     1861 aagaagttct ggaacgtgtt ccacctgcag ccgatgcgcg attataaggg cgacgtccag
     1921 tacttcattg gcgtgcagct ggatggcacc gaacgtcttc atggcgccgc tgagcgtgag
     1981 gcggtctgcc tgatcaaaaa gacagccttt cagattgctg aggcagcgaa cgacgaaaat
     2041 tactttggaa gcgggagtgg gagcaaaaag aagaaaaaga agtcaaagac aaagtgtgta
     2101 attatgtagg cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     2161 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     2221 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     2281 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     2341 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     2401 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     2461 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     2521 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     2581 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     2641 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     2701 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     2761 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     2821 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2881 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2941 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3001 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3061 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3121 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3181 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3241 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3301 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3361 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3421 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3481 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     3541 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3601 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3661 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3721 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3781 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3841 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     3901 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     3961 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4021 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4081 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4141 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4201 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4261 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     4321 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     4381 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     4441 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     4501 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     4561 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     4621 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4681 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4741 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4801 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4861 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4921 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4981 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5041 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5101 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5161 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5221 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5281 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5341 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     5401 gttatcccct gattctgtgg ataaccgtat taccgccatg
//
