LOCUS       sequence-198857-        6956 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses fusion of dynein adapter BICDN (1-594) with GFP and CIB1 
            (1-170).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6956)
  AUTHORS   Duan L, Che D, Zhang K, Ong Q, Guo S, Cui B
  TITLE     Optogenetic control of molecular motors and organelle distributions 
            in cells.
  JOURNAL   Chem Biol. 2015 May 21;22(5):671-82. doi: 
            10.1016/j.chembiol.2015.04.014. Epub 2015 May 9.
  PUBMED    25963241
REFERENCE   2  (bases 1 to 6956)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.chembiol.2015.04.014"; journalName: "Chem Biol"; date: 
            "2015-05-21- 21"; volume: "22"; issue: "5"; pages: "671-82"
COMMENT     Sequence Label: GFP-BICDN-CIBN
FEATURES             Location/Qualifiers
     source          1..6956
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     misc_feature    591..671
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      673..682
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(679..681,682..684,685..1395)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(679..681,682..684,685..1395)
                     /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(724..745)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(985..1004)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1332..1353
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3217..3726
                     /codon_start=1
                     /product="N-terminal portion of the CIB1 transcription 
                     factor from Arabidopsis thaliana"
                     /label=CIBN
                     /note="binds to CRY2 (cryptochrome 2) that has absorbed 
                     blue light (Kennedy et al., 2010)"
                     /translation="MNGAIGGDLLLNFPDMSVLERQRAHLKYLNPTFDSPLAGFFADSS
                     MITGGEMDSYLSTAGLNLPMMYGETTVEGDSRLSISPETTLGTGNFKAAKFDTETKDCN
                     EAAKKMTMNRDDLVEEGEEEKSKITEQNNGSTKSIKKMKHKAKKEENNFSNDSSKVTKE
                     LEKTDYI"
     polyA_signal    3744..3865
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3781..3800)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3835..3854
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3872..4327)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4009..4030)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4221..4240
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4354..4458
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4460..4817
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4460..4651
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4483..4503)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4668..4803
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4852..5646
                     /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             4852..5646
                     /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(4906..4925)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5516..5535
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5834..5853)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5878..5925
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6254..6842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6743..6762
                     /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 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
      661 gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
      721 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
      781 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
      841 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
      901 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      961 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     1021 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     1081 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     1141 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     1201 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     1261 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     1321 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     1381 gacgagctgt acaagtccgg ccggactcag atctcgacca tgtccgcgcc gtcggaggag
     1441 gaggagtatg cgcgtctggt gatggaggcg cagccggagt ggctgcgcgc cgaggtgaag
     1501 cggctgtccc acgagctggc cgagaccacg cgtgagaaga tccaggcggc cgagtatgga
     1561 ctggcggtgc tggaggagaa gcaccagctc aagctgcagt tcgaggagct cgaggtggac
     1621 tatgaggcca tccgcagcga gatggagcag ctcaaagagg catttggcca ggcacataca
     1681 aaccacaaga aggtggctgc tgatggtgag agccgggagg agagcctgat ccaggagtcg
     1741 gcctccaagg agcagtacta cgtgcggaag gtgctggagc tgcagacaga gctgaaacag
     1801 ctgcgcaacg tcctcaccaa cacccagtct gagaatgagc gcctcacgtc tgtggcccag
     1861 gagctaaaag agatcaacca gaatgtggag atccagcgtg gtcgcctgcg ggatgacatc
     1921 aaggagtaca agttccggga ggcccgtcta cttcaggact actcagagct ggaggaggag
     1981 aacatcagcc tgcagaaaca agtgtctgtg ctcaggcaga accaggtgga gtttgagggc
     2041 ctcaagcatg aaatcaagcg cctggaggag gagacagagt acctcaacag ccagctagag
     2101 gatgccatcc ggcttaagga gatctctgaa cggcagttgg aggaggcgct ggagacactg
     2161 aagacagagc gggagcagaa gaacaacctg cgcaaggagt tgtcgcacta catgagcatc
     2221 aacgattcct tctataccag ccacctgcag gtctccttag atggcctcaa gttcagtgat
     2281 gatactgtca ccgcagagcc caacaatgac gccgaagccc tggtcaatgg ctttgagcac
     2341 agtggcttgg tcaaatcatc cttggacaac aagacatcca cacccaggaa ggatggccta
     2401 gccccaccct cccccagcct cgtctctgac ctgctcagtg agctccacat atctgagatc
     2461 cagaagctga aacagcagct ggtgcagatg gagcgggaga aggtgggcct gctggcaaca
     2521 ctgcaggaca cacagaagca gctggagcag gctcggggaa ccctctctga gcagcatgag
     2581 aaggtgaatc gtctcacaga gaaccttagt gccctccggc gcctgcaggc tggcaaggag
     2641 cgacagacct cgttggataa tgagaaggac cgtgacagcc acgaagacgg tgactactat
     2701 gaggtggaca tcaatgggcc tgagatcctg gcctgcaagt accacgtggc tgtggctgag
     2761 gctggcgagc tccgggagca gctcaaggcg ttgcgcagca cacacgaagc tcgcgaagcc
     2821 cagcacgcag aagaaaaggg ccggtatgag gctgagggcc aggcactcac tgagaagatc
     2881 tctctgctgg agaaggctag ccaccaggac cgagagctgc tggcccatct ggaaaaggag
     2941 ctgaagaagg tgagcgatgt ggctggtgag acccagggca gcctgaatgt ggctcaggat
     3001 gagctggtga ccttcagcga ggagctggcc aacctctacc accatgtgtg catgtgcaac
     3061 aacgagacgc ccaaccgtgt catgctcgac tattatcgtg agggccaggg caaggctggc
     3121 cgcaccagcc cagagggccg cgggcgccgg tcacctgtcc tcttgcccaa ggggctgttg
     3181 gccacagagg tgggccgaga attgtacaag ggatccatga atggagctat aggaggtgac
     3241 cttttgctca attttcctga catgtcggtc ctagagcgcc aaagggctca cctcaagtac
     3301 ctcaatccca cctttgattc tcctctcgcc ggcttctttg ccgattcttc aatgattacc
     3361 ggcggcgaga tggacagcta tctttcgact gccggtttga atcttccgat gatgtacggt
     3421 gagacgacgg tggaaggtga ttcaagactc tcaatttcgc cggaaacgac gcttgggact
     3481 ggaaatttca aggcagcgaa gtttgataca gagactaagg attgtaatga ggcggcgaag
     3541 aagatgacga tgaacagaga tgacctagta gaagaaggag aagaagagaa gtcgaaaata
     3601 acagagcaaa acaatgggag cacaaaaagc atcaagaaga tgaaacacaa agccaagaaa
     3661 gaagagaaca atttctctaa tgattcatct aaagtgacga aggaattgga gaaaacggat
     3721 tatatttaac aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     3781 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3841 tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa
     3901 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     3961 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     4021 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     4081 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     4141 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     4201 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     4261 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     4321 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4381 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4441 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     4501 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4561 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4621 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4681 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4741 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4801 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     4861 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4921 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4981 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     5041 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     5101 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     5161 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     5221 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5281 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5341 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     5401 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5461 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5521 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5581 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5641 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5701 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5761 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     5821 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     5881 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     5941 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     6001 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     6061 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     6121 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6181 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6241 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6301 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6361 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6421 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6481 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6541 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6601 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6661 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6721 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6781 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6841 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6901 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
