LOCUS       sequence-369200-        6328 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Kinesin Light Chain 1 fused to GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6328)
  AUTHORS   Zamponi E, Meehl JB, Voeltz GK
  TITLE     The ER ladder is a unique morphological feature of developing 
            mammalian axons.
  JOURNAL   Dev Cell. 2022 Jun 6;57(11):1369-1382.e6. doi: 
            10.1016/j.devcel.2022.05.002. Epub 2022 May 23.
  PUBMED    35609616
REFERENCE   2  (bases 1 to 6328)
  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: 
            "10.1016/j.devcel.2022.05.002"; journalName: "Dev Cell"; date: 
            "2022-06-6- 6"; volume: "57"; issue: "11"; pages: "1369-1382.e6"
COMMENT     Sequence Label: Klc1-GFP
FEATURES             Location/Qualifiers
     source          1..6328
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2287..3006
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    3128..3249
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3165..3184)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3219..3238
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3256..3711)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3393..3414)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3605..3624
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3738..3842
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3867..3887)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3911..4121
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3933..4129
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3980..4115
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4164..4958
                     /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             4164..4958
                     /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(4218..4237)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4828..4847
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5146..5165)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5190..5237
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5566..6154
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6055..6074
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgatg
      541 tatgacaaca tgtccacaat ggtgtacata aaggaagaca agttggagaa gcttacacag
      601 gatgaaatta tttctaagac aaagcaagta attcaggggc tggaagcttt gaagaatgag
      661 cacaattcca ttttacaaag tttgctggag acactgaagt gtttgaagaa agatgatgaa
      721 agtaatttgg tggaggagaa atcaaacatg atccggaagt cactggagat gttggagctc
      781 ggcctgagtg aggcacaggt tatgatggct ttgtcaaatc acctgaatgc tgtggagtcc
      841 gagaagcaga aactgcgtgc gcaggttcgt cgtctgtgcc aggagaatca gtggctacgg
      901 gatgaactgg ccaacacgca gcagaaactg cagaagagtg agcagtctgt ggctcaactg
      961 gaggaggaga agaagcatct ggagtttatg aatcagctaa aaaaatatga tgacgacatt
     1021 tccccatccg aggacaaaga cactgattct accaaagagc ctctggatga ccttttcccc
     1081 aatgatgaag acgacccagg gcaaggaatc cagcagcagc acagcagtgc agccgcggct
     1141 gcccagcagg gcggctacga gatccccgcg cggctgcgga cgctccacaa cctggtgatc
     1201 cagtacgcct cgcaggggcg ctacgaggta gctgtgcccc tctgcaagca ggccctggag
     1261 gacctggaga agacttcagg acacgaccac ccggacgtgg ccaccatgct caacatcctg
     1321 gccttggtgt acagggatca gaataaatac aaagatgcag ctaacctact gaatgatgcc
     1381 ttggctattc gtgagaaaac tttgggcaaa gatcatcctg cggtggcggc gactttgaat
     1441 aaccttgcag tcctttatgg taaaagaggg aagtacaaag aagcagagcc gttgtgtaaa
     1501 agagctctgg aaatccgaga aaaggttttg gggaaggatc accccgatgt tgccaagcag
     1561 ttaaataact tggccttact gtgccagaac cagggcaagt atgaagaagt agaatattat
     1621 tatcaaagag ccctcgagat ctaccagaca aaactgggac ctgatgaccc caacgtggct
     1681 aagacgaaaa ataacctggc atcctgctat ttgaaacaag gaaagttcaa gcaagcagaa
     1741 acactgtaca aagagattct cactcgtgca catgaaaggg agtttggttc tgtagatgat
     1801 gaaaataaac ccatctggat gcatgctgaa gaaagagaag aatgcaaagg aaagcaaaag
     1861 gatgggacat cttttggaga gtatggcggc tggtacaaag cctgcaaagt tgatagtcca
     1921 actgttacaa ccactctaaa aaaccttggg gcactttaca gacgtcaagg caaatttgaa
     1981 gctgcagaaa cgttagaaga agctgctatg aggtctcgta aacagggtct tgacaatgtt
     2041 cacaaacaga gggtggcaga agtgctcaat gaccctgaga acatggagaa gcgcaggagc
     2101 cgtgagagcc tcaacgtgga cgtggtcaag tacgagagtg gccctgacgg aggggaggaa
     2161 gtgagtatga gcgtagagtg gaacgggggc gtctctggcc gagcctcttt ttgtggaaaa
     2221 cgacagcagc agcagtggcc tggaagacgc caccgccggg taccgcgggc ccgggatcca
     2281 ccggtcatgg tgagcaaggg cgccgagctg ttcaccggca tcgtgcccat cctgatcgag
     2341 ctgaatggcg atgtgaatgg ccacaagttc agcgtgagcg gcgagggcga gggcgatgcc
     2401 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc tgtgccctgg
     2461 cccaccctgg tgaccaccct gagctacggc gtgcagtgct tctcacgcta ccccgatcac
     2521 atgaagcagc acgacttctt caagagcgcc atgcctgagg gctacatcca ggagcgcacc
     2581 atcttcttcg aggatgacgg caactacaag tcgcgcgccg aggtgaagtt cgagggcgat
     2641 accctggtga atcgcatcga gctgaccggc accgatttca aggaggatgg caacatcctg
     2701 ggcaataaga tggagtacaa ctacaacgcc cacaatgtgt acatcatgac cgacaaggcc
     2761 aagaatggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggatgg cagcgtgcag
     2821 ctggccgacc actaccagca gaataccccc atcggcgatg gccctgtgct gctgcccgat
     2881 aaccactacc tgtccaccca gagcgccctg tccaaggacc ccaacgagaa gcgcgatcac
     2941 atgatctact tcggcttcgt gaccgccgcc gccatcaccc acggcatgga tgagctgtac
     3001 aagtgagcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     3061 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     3121 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3181 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3241 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     3301 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     3361 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     3421 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     3481 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3541 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3601 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3661 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     3721 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3781 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3841 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3901 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3961 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4021 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4081 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     4141 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4201 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4261 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4321 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     4381 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4441 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4501 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4561 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4621 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4681 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4741 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4801 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4861 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4921 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4981 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5041 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5101 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     5161 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     5221 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     5281 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     5341 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     5401 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5461 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5521 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5581 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5641 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5701 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5761 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5821 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5881 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5941 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6001 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6061 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6121 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6181 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6241 tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat
     6301 tagttcatag cccatatatg gagttccg
//
