LOCUS       sequence-438070-        9071 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Halo7 fused to alpha tubulin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9071)
  TITLE     Janelia Integrative Imaging
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9071)
  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            "Unpublished"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: Halo_aTubulin-pTwistCMVBetaGlobWPRENeo
FEATURES             Location/Qualifiers
     source          1..9071
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             5..892
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     misc_feature    2351..2939
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2404..2424)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2822..2833)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2992..3113
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3003..3022)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3057..3076
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3228..3274
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3304..3440
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3485..3556
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     promoter        3601..3797
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3648..3783
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3811..4605
                     /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(3865..3884)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4475..4494
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4616..4840
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(5036..5624)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5116..5135)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        6370..6474
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6475..6543,6544..7335)
                     /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(6693..6712)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7348..7484
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7629..7700
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7705..7751)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7769..7817
                     /note="synthetic polyadenylation signal"
     enhancer        7898..8201
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8203..8405
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8356..8376
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          8539..9014
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
     regulatory      join(9070..9071,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggaa atcggtactg gctttccatt cgacccccat tatgtggaag tcctgggcga
       61 gcgcatgcac tacgtcgatg ttggtccgcg cgatggcacc cctgtgctgt tcctgcacgg
      121 taacccgacc tcctcctacg tgtggcgcaa catcatcccg catgttgcac cgacccatcg
      181 ctgcattgct ccagacctga tcggtatggg caaatccgac aaaccagacc tgggttattt
      241 cttcgacgac cacgtccgct tcatggatgc cttcatcgaa gccctgggtc tggaagaggt
      301 cgtcctggtc attcacgact ggggctccgc tctgggtttc cactgggcca agcgcaatcc
      361 agagcgcgtc aaaggtattg catttatgga gttcatccgc cctatcccga cctgggacga
      421 atggccagaa tttgcccgcg agaccttcca ggccttccgc accaccgacg tcggccgcaa
      481 gctgatcatc gatcagaacg tttttatcga gggtacgctg ccgatgggtg tcgtccgccc
      541 gctgactgaa gtcgagatgg accattaccg cgagccgttc ctgaatcctg ttgaccgcga
      601 gccactgtgg cgcttcccaa acgagctgcc aatcgccggt gagccagcga acatcgtcgc
      661 gctggtcgaa gaatacatgg actggctgca ccagtcccct gtcccgaagc tgctgttctg
      721 gggcacccca ggcgttctga tcccaccggc cgaagccgct cgcctggcca aaagcctgcc
      781 taactgcaag gctgtggaca tcggcccggg tctgaatctg ctgcaagaag acaacccgga
      841 cctgatcggc agcgagatcg cgcgctggct gtctactctg gagatttccg gtggagcatc
      901 gggagcctca ggagcatcgc tcagatctcg agtgcgtgag tgcatctcca tccacgttgg
      961 ccaggctggt gtccagattg gcaatgcctg ctgggagctc tactgcctgg aacacggcat
     1021 ccagcccgat ggccagatgc caagtgacaa gaccattggg ggaggagatg actccttcaa
     1081 caccttcttc agtgagacgg gcgctggcaa gcacgtgccc cgggctgtgt ttgtagactt
     1141 ggaacccaca gtcattgatg aagttcgcac tggcacctac cgccagctct tccaccctga
     1201 gcagctcatc acaggcaagg aagatgctgc caataactat gcccgagggc actacaccat
     1261 tggcaaggag atcattgacc ttgtgttgga ccgaattcgc aagctggctg accagtgcac
     1321 cggtcttcag ggcttcttgg ttttccacag ctttggtggg ggaactggtt ctgggttcac
     1381 ctccctgctc atggaacgtc tctcagttga ttatggcaag aagtccaagc tggagttctc
     1441 catttaccca gcaccccagg tttccacagc tgtagttgag ccctacaact ccatcctcac
     1501 cacccacacc accctggagc actctgattg tgccttcatg gtagacaatg aggccatcta
     1561 tgacatctgt cgtagaaacc tcgatatcga gcgcccaacc tacactaacc ttaaccgcct
     1621 tattagccag attgtgtcct ccatcactgc ttccctgaga tttgatggag ccctgaatgt
     1681 tgacctgaca gaattccaga ccaacctggt gccctacccc cgcatccact tccctctggc
     1741 cacatatgcc cctgtcatct ctgctgagaa agcctaccat gaacagcttt ctgtagcaga
     1801 gatcaccaat gcttgctttg agccagccaa ccagatggtg aaatgtgacc ctcgccatgg
     1861 taaatacatg gcttgctgcc tgttgtaccg tggtgacgtg gttcccaaag atgtcaatgc
     1921 tgccattgcc accatcaaaa ccaagcgcag catccagttt gtggattggt gccccactgg
     1981 cttcaaggtt ggcatcaact accagcctcc cactgtggtg cctggtggag acctggccaa
     2041 ggtacagaga gctgtgtgca tgctgagcaa caccacagcc attgctgagg cctgggctcg
     2101 cctggaccac aagtttgacc tgatgtatgc caagcgtgcc tttgttcact ggtacgtggg
     2161 tgaggggatg gaggaaggcg agttttcaga ggcccgtgaa gatatggctg cccttgagaa
     2221 ggattatgag gaggttggtg tggattctgt tgaaggagag ggtgaggaag aaggagagga
     2281 atactaatct agaagttgtc tcctcctgca ctgactgact gatacaatcg atttctggat
     2341 ccgcaggcct aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2401 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2461 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     2521 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     2581 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     2641 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     2701 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     2761 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     2821 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     2881 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     2941 tgtctagctt gactgactga gatacagcgt accttcagct cacagacatg ataagataca
     3001 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     3061 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     3121 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     3181 agtaaaacct ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct
     3241 tggggcctct aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta
     3301 ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata
     3361 tgccagattc agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct
     3421 taccagaaat ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc
     3481 aagacattcc tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa
     3541 ctttcttccc tccctaatct cattggttac cttgggctat cgaaacttaa ttaagcgatc
     3601 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3661 ctccgcccag ttccgcccat tctccgcccc atcgctgact aatttttttt atttatgcag
     3721 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3781 gcctaggctt ttgcaaagga ggtagccaac atgattgaac aagatggatt gcacgcaggt
     3841 tctcccgccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3901 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3961 accgacctgt ccggtgccct gaatgaactc caggacgagg cagcgcggct atcgtggctg
     4021 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4081 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4141 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4201 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4261 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4321 ttcgccaggc tcaaggcgcg gatgcccgac ggcgaggatc tcgtcgtgac ccacggcgat
     4381 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4441 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4501 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4561 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctagtatgt aagccctgtg
     4621 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     4681 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     4741 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     4801 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg ttaattaacc agtcaagtca
     4861 gctacttggc gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag
     4921 ttcgatctgg tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt
     4981 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5041 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5101 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5161 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5221 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5281 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5341 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5401 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5461 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5521 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5581 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5641 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5701 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     5761 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     5821 ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc
     5881 caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct
     5941 cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc
     6001 ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt
     6061 caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg
     6121 cctggtgtac cgagaacgat cctctcagtg cgagtctcga cgatccatat cgttgcttgg
     6181 cagtcagcca gtcggaatcc agcttgggac ccaggaagtc caatcgtcag atattgtact
     6241 caagcctggt cacggcagcg taccgatctg tttaaaccta gatattgata gtctgatcgg
     6301 tcaacgtata atcgagtcct agcttttgca aacatctatc aagagacagg atcagcagga
     6361 ggctttcgcc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     6421 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     6481 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     6541 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg
     6601 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     6661 cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     6721 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     6781 tattcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     6841 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga
     6901 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6961 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     7021 gcaatggcaa caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg
     7081 caacagttga tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     7141 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     7201 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     7261 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     7321 attaagcatt ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg
     7381 ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct tccccaactt
     7441 gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaagatcgt ttaaactcga
     7501 ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt
     7561 gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg
     7621 gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac
     7681 agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag
     7741 gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga
     7801 tggatagcga ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta
     7861 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     7921 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     7981 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     8041 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     8101 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     8161 cttggcagta catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt
     8221 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     8281 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     8341 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     8401 gagctggttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc
     8461 atagaagaca ccgggaccga tccagcctcc cctcgaagct tacatgtggt accgagctcg
     8521 gatcctgaga acttcagggt gagtctatgg gacccttgat gttttctttc cccttctttt
     8581 ctatggttaa gttcatgtca taggaagggg agaagtaaca gggtacacat attgaccaaa
     8641 tcagggtaat tttgcatttg taattttaaa aaatgctttc ttcttttaat atactttttt
     8701 gtttatctta tttctaatac tttccctaat ctctttcttt cagggcaata atgatacaat
     8761 gtatcatgcc tctttgcacc attctaaaga ataacagtga taatttctgg gttaaggcaa
     8821 tagcaatatt tctgcatata aatatttctg catataaatt gtaactgatg taagaggttt
     8881 catattgcta atagcagcta caatccagct accattctgc ttttatttta tggttgggat
     8941 aaggctggat tattctgagt ccaagctagg cccttttgct aatcatgttc atacctctta
     9001 tcttcctccc acagctcctg ggcaacgtgc tggtctgtgt gctggcccat cactttggca
     9061 aaggcggccg c
//
