LOCUS       sequence-438068-        7733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Halo7 targeted to the plasma membrane.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7733)
  TITLE     Janelia Integrative Imaging
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7733)
  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            "Unpublished"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: Lck-Halo_pTwistCMVBetaGlobWPRENeo
FEATURES             Location/Qualifiers
     source          1..7733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             62..946
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     misc_feature    1013..1601
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1066..1086)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1484..1495)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1654..1775
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1665..1684)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1719..1738
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1890..1936
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1966..2102
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2147..2218
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     promoter        2263..2459
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2310..2445
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2473..3267
                     /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(2527..2546)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3137..3156
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3278..3502
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3698..4286)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3778..3797)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5032..5136
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5137..5205,5206..5997)
                     /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(5355..5374)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6010..6146
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6291..6362
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6367..6413)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6431..6479
                     /note="synthetic polyadenylation signal"
     enhancer        6560..6863
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6865..7067
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7018..7038
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          7201..7676
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
     regulatory      join(7732..7733,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatgggc tgcgtgtgca gcagcaaccc cgagggagca tcgggagcct caggagcatc
       61 ggaaatcggt actggctttc cattcgaccc ccattatgtg gaagtcctgg gcgagcgcat
      121 gcactacgtc gatgttggtc cgcgcgatgg cacccctgtg ctgttcctgc acggtaaccc
      181 gacctcctcc tacgtgtggc gcaacatcat cccgcatgtt gcaccgaccc atcgctgcat
      241 tgctccagac ctgatcggta tgggcaaatc cgacaaacca gacctgggtt atttcttcga
      301 cgaccacgtc cgcttcatgg atgccttcat cgaagccctg ggtctggaag aggtcgtcct
      361 ggtcattcac gactggggct ccgctctggg tttccactgg gccaagcgca atccagagcg
      421 cgtcaaaggt attgcattta tggagttcat ccgccctatc ccgacctggg acgaatggcc
      481 agaatttgcc cgcgagacct tccaggcctt ccgcaccacc gacgtcggcc gcaagctgat
      541 catcgatcag aacgttttta tcgagggtac gctgccgatg ggtgtcgtcc gcccgctgac
      601 tgaagtcgag atggaccatt accgcgagcc gttcctgaat cctgttgacc gcgagccact
      661 gtggcgcttc ccaaacgagc tgccaatcgc cggtgagcca gcgaacatcg tcgcgctggt
      721 cgaagaatac atggactggc tgcaccagtc ccctgtcccg aagctgctgt tctggggcac
      781 cccaggcgtt ctgatcccac cggccgaagc cgctcgcctg gccaaaagcc tgcctaactg
      841 caaggctgtg gacatcggcc cgggtctgaa tctgctgcaa gaagacaacc cggacctgat
      901 cggcagcgag atcgcgcgct ggctgtctac tctggagatt tccggttaat ctagaagttg
      961 tctcctcctg cactgactga ctgatacaat cgatttctgg atccgcaggc ctaatcaacc
     1021 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     1081 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     1141 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     1201 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     1261 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     1321 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     1381 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     1441 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     1501 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     1561 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctgtctagc ttgactgact
     1621 gagatacagc gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa
     1681 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     1741 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     1801 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     1861 gtggtattgg cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt
     1921 cttgaggggt tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa
     1981 aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg
     2041 atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt
     2101 aaggtcgtca gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg
     2161 tcttttctgg acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat
     2221 ctcattggtt accttgggct atcgaaactt aattaagcga tctgcatctc aattagtcag
     2281 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     2341 attctccgcc ccatcgctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     2401 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     2461 gaggtagcca acatgattga acaagatgga ttgcacgcag gttctcccgc cgcttgggtg
     2521 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     2581 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     2641 ctgaatgaac tccaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     2701 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     2761 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     2821 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     2881 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     2941 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     3001 cggatgcccg acggcgagga tctcgtcgtg acccacggcg atgcctgctt gccgaatatc
     3061 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     3121 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     3181 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     3241 tatcgccttc ttgacgagtt cttctagtat gtaagccctg tgccttctag ttgccagcca
     3301 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3361 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3421 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     3481 ggggatgcgg tgggctctat ggttaattaa ccagtcaagt cagctacttg gcgagatcga
     3541 cttgtctggg tttcgactac gctcagaatt gcgtcagtca agttcgatct ggtccttgct
     3601 attgcacccg ttctccgatt acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa
     3661 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3721 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3781 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3841 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3901 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3961 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4021 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4081 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4141 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4201 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4261 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4321 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4381 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4441 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4501 tatttcgttc atccatagtt gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa
     4561 atcttcaaac ctttcgtccg atccatcttg caggctacct ctcgaacgaa ctatcgcaag
     4621 tctcttggcc ggccttgcgc cttggctatt gcttggcagc gcctatcgcc aggtattact
     4681 ccaatcccga atatccgaga tcgggatcac ccgagagaag ttcaacctac atcctcaatc
     4741 ccgatctatc cgagatccga ggaatatcga aatcggggcg cgcctggtgt accgagaacg
     4801 atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat
     4861 ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag
     4921 cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc
     4981 ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg ccgcggaacc
     5041 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5101 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5161 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5221 gtgaaagtaa aagatgctga agatcagttg ggtgcrcgag tgggttacat cgaactggat
     5281 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgctttcc aatgatgagc
     5341 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5401 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtattcacc agtcacagaa
     5461 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5521 gataacactg cggccaactt acttctgaca acgattggag gaccgaagga gctaaccgct
     5581 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5641 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaaccttg
     5701 cgtaaactat taactggcga actacttact ctagcttccc ggcaacagtt gatagactgg
     5761 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5821 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5881 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5941 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaaccg
     6001 attctaggtg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc
     6061 cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt
     6121 cctccttacc agaaatttat ccttaagatc gtttaaactc gactctggct ctatcgaatc
     6181 tccgtcgttt cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat
     6241 ctcgtcagct atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac
     6301 cattagctcc acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc
     6361 tcaattcaaa aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg
     6421 ttgcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta
     6481 tctaactgct gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata
     6541 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     6601 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     6661 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     6721 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     6781 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     6841 tattagtcat cgctattacc atgctgatgc ggttttggca gtacatcaat gggcgtggat
     6901 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     6961 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     7021 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
     7081 gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc
     7141 gatccagcct cccctcgaag cttacatgtg gtaccgagct cggatcctga gaacttcagg
     7201 gtgagtctat gggacccttg atgttttctt tccccttctt ttctatggtt aagttcatgt
     7261 cataggaagg ggagaagtaa cagggtacac atattgacca aatcagggta attttgcatt
     7321 tgtaatttta aaaaatgctt tcttctttta atatactttt ttgtttatct tatttctaat
     7381 actttcccta atctctttct ttcagggcaa taatgataca atgtatcatg cctctttgca
     7441 ccattctaaa gaataacagt gataatttct gggttaaggc aatagcaata tttctgcata
     7501 taaatatttc tgcatataaa ttgtaactga tgtaagaggt ttcatattgc taatagcagc
     7561 tacaatccag ctaccattct gcttttattt tatggttggg ataaggctgg attattctga
     7621 gtccaagcta ggcccttttg ctaatcatgt tcatacctct tatcttcctc ccacagctcc
     7681 tgggcaacgt gctggtctgt gtgctggccc atcactttgg caaaggcggc cgc
//
