LOCUS       sequence-368223-        6045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization of LSSmGFP fluorescent protein (ex/em = 400/510 nm) to 
            tubulin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6045)
  AUTHORS   Campbell BC, Paez-Segala MG, Looger LL, Petsko GA, Liu CF
  TITLE     Chemically stable fluorescent proteins for advanced microscopy.
  JOURNAL   Nat Methods. 2022 Nov 7. pii: 10.1038/s41592-022-01660-7. doi: 
            10.1038/s41592-022-01660-7.
  PUBMED    36344833
REFERENCE   2  (bases 1 to 6045)
  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"; journalName: "Nat 
            Methods. 2022 Nov 7. pii: 10.1038/s41592-022-01660-7. doi: 
            10.1038/s41592-022-01660-7."
COMMENT     Sequence Label: Tubulin-LSSmGFP
FEATURES             Location/Qualifiers
     source          1..6045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        149..352
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     303..323
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      391..400
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(442..463)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    2617..2738
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2654..2673)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2708..2727
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2745..3200)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2882..2903)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3094..3113
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3227..3331
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3333..3690
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3333..3524
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3356..3376)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3541..3676
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3725..4519
                     /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             3725..4519
                     /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(3779..3798)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4389..4408
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4707..4726)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4751..4798
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5127..5715
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5616..5635
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        join(5890..6045,1..148)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
       61 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      121 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      181 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      241 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      301 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
      361 gaaccgtcag atccgctagc gctaccggtc gccaccatgg tgagcaaggg cgaggagctg
      421 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
      481 agcgtccgcg gcgagggcga gggcgatgcc accaacggca aactgtccct gaagctcatc
      541 tccaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt aagctacggc
      601 cagatggtgt tcgcccgcta ccctgacaac atgaagcagc acgacttctt caagtccgcc
      661 atgcccgaag gctacgtcca ggagcgcacc atctctttcg aggacgacgg ttactacaag
      721 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcgt gctgaagggc
      781 atcgacttca aggaggacgg caacatcctg gggcacaacc tggagtacaa cttcaacagc
      841 cacaacgtct atatcacggc cgacaagcag aagaacggca tcaaggctaa cttcaagatc
      901 cgccacaacg ttgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc
      961 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcatcca gtccaagctg
     1021 agcaaagacc ccaacgagaa gcgcgatcac atggtcctga aggagagggt gaccgccgcc
     1081 gggattacac atgacatgaa cgagctgtac aagtccggac tcagatctcg agtgcgtgag
     1141 tgcatctcca tccacgttgg ccaggctggt gtccagattg gcaatgcctg ctgggagctc
     1201 tactgcctgg aacacggcat ccagcccgat ggccagatgc caagtgacaa gaccattggg
     1261 ggaggagatg actccttcaa caccttcttc agtgagacgg gcgctggcaa gcacgtgccc
     1321 cgggctgtgt ttgtagactt ggaacccaca gtcattgatg aagttcgcac tggcacctac
     1381 cgccagctct tccaccctga gcagctcatc acaggcaagg aagatgctgc caataactat
     1441 gcccgagggc actacaccat tggcaaggag atcattgacc ttgtgttgga ccgaattcgc
     1501 aagctggctg accagtgcac cggtcttcag ggcttcttgg ttttccacag ctttggtggg
     1561 ggaactggtt ctgggttcac ctccctgctc atggaacgtc tctcagttga ttatggcaag
     1621 aagtccaagc tggagttctc catttaccca gcaccccagg tttccacagc tgtagttgag
     1681 ccctacaact ccatcctcac cacccacacc accctggagc actctgattg tgccttcatg
     1741 gtagacaatg aggccatcta tgacatctgt cgtagaaacc tcgatatcga gcgcccaacc
     1801 tacactaacc ttaaccgcct tattagccag attgtgtcct ccatcactgc ttccctgaga
     1861 tttgatggag ccctgaatgt tgacctgaca gaattccaga ccaacctggt gccctacccc
     1921 cgcatccact tccctctggc cacatatgcc cctgtcatct ctgctgagaa agcctaccat
     1981 gaacagcttt ctgtagcaga gatcaccaat gcttgctttg agccagccaa ccagatggtg
     2041 aaatgtgacc ctcgccatgg taaatacatg gcttgctgcc tgttgtaccg tggtgacgtg
     2101 gttcccaaag atgtcaatgc tgccattgcc accatcaaaa ccaagcgcag catccagttt
     2161 gtggattggt gccccactgg cttcaaggtt ggcatcaact accagcctcc cactgtggtg
     2221 cctggtggag acctggccaa ggtacagaga gctgtgtgca tgctgagcaa caccacagcc
     2281 attgctgagg cctgggctcg cctggaccac aagtttgacc tgatgtatgc caagcgtgcc
     2341 tttgttcact ggtacgtggg tgaggggatg gaggaaggcg agttttcaga ggcccgtgaa
     2401 gatatggctg cccttgagaa ggattatgag gaggttggtg tggattctgt tgaaggagag
     2461 ggtgaggaag aaggagagga atactaagga tccaccggat ctagataact gatcataatc
     2521 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     2581 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     2641 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2701 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa
     2761 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     2821 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     2881 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     2941 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     3001 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     3061 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     3121 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     3181 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3241 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3301 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     3361 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3421 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3481 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3541 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3601 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     3661 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     3721 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3781 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3841 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3901 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3961 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4021 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4081 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4141 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4201 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     4261 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4321 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4381 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4441 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4501 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4561 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4621 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4681 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     4741 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     4801 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     4861 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     4921 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     4981 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5041 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5101 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5161 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5221 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5281 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5341 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5401 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5461 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5521 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5581 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5641 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5701 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5761 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5821 gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     5881 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     5941 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     6001 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggc
//
