LOCUS       sequence-438071-        8891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses alpha tubulin fused to mStayGold.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8891)
  TITLE     Janelia Integrative Imaging
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8891)
  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: 
            "Unpublished"
COMMENT     Sequence Label: mStayGold-aTubulin_pTwistCMVBetaGlobWPRENeo
FEATURES             Location/Qualifiers
     source          1..8891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             5..682
                     /codon_start=1
                     /product="bright and highly photostable monomeric green 
                     fluorescent protein (Ando et al., 2023)"
                     /label=mStayGold(J)
                     /note="includes multiple mutations plus an N-terminal 
                     iinker sequence"
                     /translation="MVSTGEELFTGVVPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKG
                     KYVCTSGKLPMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQYKGDGS
                     IHAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTGDMDVSLPNEVQHIPIDDGVECTVTL
                     QYPLLSDESKCVEAYQNTIIKPLHNQPAPDVPFHWIRKQYTQSKDDTEERDHIIQSETL
                     EAHL"
     misc_feature    2171..2759
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2224..2244)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2642..2653)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2812..2933
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2823..2842)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2877..2896
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3048..3094
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3124..3260
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3305..3376
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     promoter        3421..3617
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3468..3603
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3631..4425
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3685..3704)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4295..4314
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4436..4660
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(4856..5444)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4936..4955)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        6190..6294
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6295..6363,6364..7155)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6513..6532)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7168..7304
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7449..7520
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7525..7571)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7589..7637
                     /note="synthetic polyadenylation signal"
     enhancer        7718..8021
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8023..8225
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8176..8196
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          8359..8834
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
     regulatory      join(8890..8891,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggtg tctacaggcg aggagctgtt taccggcgtg gtgcccttca agttccagct
       61 gaagggcacc atcaacggca agagcttcac cgtggaaggg gaaggtgaag ggaacagcca
      121 cgagggcagc cacaaaggca agtacgtgtg caccagcggc aaactgccaa tgtcttgggc
      181 cgccctggga actagcttcg gctatggcat gaagtactac accaagtacc ccagcggcct
      241 gaagaactgg ttccacgagg tgatgccgga agggtttact tacgacagac acatccagta
      301 caagggcgac ggcagcatcc acgccaagca ccagcacttc atgaagaacg gcacctacca
      361 caacatcgtg gagttcaccg gccaggactt caaggagaac agccccgtgc tgaccggcga
      421 catggacgtg agcctgccca acgaggtgca gcacatcccc atagatgacg gcgtggagtg
      481 cacagtgacc ctgcagtacc ctctgctgag cgacgagagc aagtgcgtgg aggcctacca
      541 gaacaccatc atcaagcccc tgcacaatca gccagccccc gatgtgccat tccactggat
      601 cagaaagcag tacacccaga gcaaggacga caccgaggag agagaccaca tcatccagag
      661 cgagaccctg gaggcccacc tgccatggca cgagccttct gcttctgccg tgggagcatc
      721 gggagcctca ggagcatcgc tcagatctcg agtgcgtgag tgcatctcca tccacgttgg
      781 ccaggctggt gtccagattg gcaatgcctg ctgggagctc tactgcctgg aacacggcat
      841 ccagcccgat ggccagatgc caagtgacaa gaccattggg ggaggagatg actccttcaa
      901 caccttcttc agtgagacgg gcgctggcaa gcacgtgccc cgggctgtgt ttgtagactt
      961 ggaacccaca gtcattgatg aagttcgcac tggcacctac cgccagctct tccaccctga
     1021 gcagctcatc acaggcaagg aagatgctgc caataactat gcccgagggc actacaccat
     1081 tggcaaggag atcattgacc ttgtgttgga ccgaattcgc aagctggctg accagtgcac
     1141 cggtcttcag ggcttcttgg ttttccacag ctttggtggg ggaactggtt ctgggttcac
     1201 ctccctgctc atggaacgtc tctcagttga ttatggcaag aagtccaagc tggagttctc
     1261 catttaccca gcaccccagg tttccacagc tgtagttgag ccctacaact ccatcctcac
     1321 cacccacacc accctggagc actctgattg tgccttcatg gtagacaatg aggccatcta
     1381 tgacatctgt cgtagaaacc tcgatatcga gcgcccaacc tacactaacc ttaaccgcct
     1441 tattagccag attgtgtcct ccatcactgc ttccctgaga tttgatggag ccctgaatgt
     1501 tgacctgaca gaattccaga ccaacctggt gccctacccc cgcatccact tccctctggc
     1561 cacatatgcc cctgtcatct ctgctgagaa agcctaccat gaacagcttt ctgtagcaga
     1621 gatcaccaat gcttgctttg agccagccaa ccagatggtg aaatgtgacc ctcgccatgg
     1681 taaatacatg gcttgctgcc tgttgtaccg tggtgacgtg gttcccaaag atgtcaatgc
     1741 tgccattgcc accatcaaaa ccaagcgcag catccagttt gtggattggt gccccactgg
     1801 cttcaaggtt ggcatcaact accagcctcc cactgtggtg cctggtggag acctggccaa
     1861 ggtacagaga gctgtgtgca tgctgagcaa caccacagcc attgctgagg cctgggctcg
     1921 cctggaccac aagtttgacc tgatgtatgc caagcgtgcc tttgttcact ggtacgtggg
     1981 tgaggggatg gaggaaggcg agttttcaga ggcccgtgaa gatatggctg cccttgagaa
     2041 ggattatgag gaggttggtg tggattctgt tgaaggagag ggtgaggaag aaggagagga
     2101 atactaatct agaagttgtc tcctcctgca ctgactgact gatacaatcg atttctggat
     2161 ccgcaggcct aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2221 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2281 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     2341 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     2401 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     2461 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     2521 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     2581 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     2641 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     2701 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     2761 tgtctagctt gactgactga gatacagcgt accttcagct cacagacatg ataagataca
     2821 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     2881 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     2941 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     3001 agtaaaacct ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct
     3061 tggggcctct aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta
     3121 ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata
     3181 tgccagattc agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct
     3241 taccagaaat ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc
     3301 aagacattcc tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa
     3361 ctttcttccc tccctaatct cattggttac cttgggctat cgaaacttaa ttaagcgatc
     3421 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3481 ctccgcccag ttccgcccat tctccgcccc atcgctgact aatttttttt atttatgcag
     3541 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3601 gcctaggctt ttgcaaagga ggtagccaac atgattgaac aagatggatt gcacgcaggt
     3661 tctcccgccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3721 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3781 accgacctgt ccggtgccct gaatgaactc caggacgagg cagcgcggct atcgtggctg
     3841 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3901 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3961 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4021 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4081 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4141 ttcgccaggc tcaaggcgcg gatgcccgac ggcgaggatc tcgtcgtgac ccacggcgat
     4201 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4261 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4321 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4381 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctagtatgt aagccctgtg
     4441 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     4501 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     4561 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     4621 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg ttaattaacc agtcaagtca
     4681 gctacttggc gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag
     4741 ttcgatctgg tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt
     4801 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4861 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4921 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4981 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5041 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5101 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5161 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5221 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5281 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5341 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5401 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5461 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5521 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     5581 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     5641 ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc
     5701 caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct
     5761 cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc
     5821 ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt
     5881 caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg
     5941 cctggtgtac cgagaacgat cctctcagtg cgagtctcga cgatccatat cgttgcttgg
     6001 cagtcagcca gtcggaatcc agcttgggac ccaggaagtc caatcgtcag atattgtact
     6061 caagcctggt cacggcagcg taccgatctg tttaaaccta gatattgata gtctgatcgg
     6121 tcaacgtata atcgagtcct agcttttgca aacatctatc aagagacagg atcagcagga
     6181 ggctttcgcc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     6241 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     6301 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     6361 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg
     6421 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     6481 cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     6541 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     6601 tattcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     6661 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga
     6721 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6781 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     6841 gcaatggcaa caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg
     6901 caacagttga tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     6961 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     7021 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     7081 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     7141 attaagcatt ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg
     7201 ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct tccccaactt
     7261 gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaagatcgt ttaaactcga
     7321 ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt
     7381 gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg
     7441 gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac
     7501 agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag
     7561 gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga
     7621 tggatagcga ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta
     7681 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     7741 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     7801 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     7861 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     7921 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     7981 cttggcagta catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt
     8041 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     8101 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     8161 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     8221 gagctggttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc
     8281 atagaagaca ccgggaccga tccagcctcc cctcgaagct tacatgtggt accgagctcg
     8341 gatcctgaga acttcagggt gagtctatgg gacccttgat gttttctttc cccttctttt
     8401 ctatggttaa gttcatgtca taggaagggg agaagtaaca gggtacacat attgaccaaa
     8461 tcagggtaat tttgcatttg taattttaaa aaatgctttc ttcttttaat atactttttt
     8521 gtttatctta tttctaatac tttccctaat ctctttcttt cagggcaata atgatacaat
     8581 gtatcatgcc tctttgcacc attctaaaga ataacagtga taatttctgg gttaaggcaa
     8641 tagcaatatt tctgcatata aatatttctg catataaatt gtaactgatg taagaggttt
     8701 catattgcta atagcagcta caatccagct accattctgc ttttatttta tggttgggat
     8761 aaggctggat tattctgagt ccaagctagg cccttttgct aatcatgttc atacctctta
     8821 tcttcctccc acagctcctg ggcaacgtgc tggtctgtgt gctggcccat cactttggca
     8881 aaggcggccg c
//
