LOCUS       sequence-369501-        5696 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses fluorescent HaloTag7-mTurquoise2 fusion protein in 
            mammalian cells with localization in the endoplasmic reticulum..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5696)
  TITLE     Organelle-specific, mTurquoise2-HaloTag fusions
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5696)
  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            "Unpublished"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: HT7-mTurquoise2-KDEL
FEATURES             Location/Qualifiers
     source          1..5696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        133..436
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        437..640
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     591..611
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     sig_peptide     713..766
                     /codon_start=1
                     /product="signal sequence of chicken lysozyme"
                     /label=lysozyme signal sequence
                     /translation="MRSLLILVLCFLPLAALG"
     regulatory      815..824
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             821..1708
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     CDS             join(1709..1711,1712..2422)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1751..1772)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2012..2031)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2359..2380
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2556..2677
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2593..2612)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2647..2666
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2684..3139)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2821..2842)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3033..3052
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3166..3270
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3272..3629
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3272..3463
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3295..3315)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3480..3615
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3664..4458
                     /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             3664..4458
                     /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(3718..3737)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4328..4347
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4646..4665)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4690..4737
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5066..5654
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5555..5574
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
       61 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      121 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      181 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      241 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      301 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      361 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      421 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      481 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      541 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      601 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
      661 ccgctagcgc taccggactc agatctcgag ctcaagcttc gaattcgcca ccatgaggtc
      721 tttgctaatc ttggtgcttt gcttcctgcc cctggctgct ctggggaaag tctttggacg
      781 atgtgagctg gcagcggcta tgaagccacc ggtcgccacc atggaaatcg gtactggctt
      841 tccattcgac ccccattatg tggaagtcct gggcgagcgc atgcactacg tcgatgttgg
      901 tccgcgcgat ggcacccctg tgctgttcct gcacggtaac ccgacctcct cctacgtgtg
      961 gcgcaacatc atcccgcatg ttgcaccgac ccatcgctgc attgctccag acctgatcgg
     1021 tatgggcaaa tccgacaaac cagacctggg ttatttcttc gacgaccacg tccgcttcat
     1081 ggatgccttc atcgaagccc tgggtctgga agaggtcgtc ctggtcattc acgactgggg
     1141 ctccgctctg ggtttccact gggccaagcg caatccagag cgcgtcaaag gtattgcatt
     1201 tatggagttc atccgcccta tcccgacctg ggacgaatgg ccagaatttg cccgcgagac
     1261 cttccaggcc ttccgcacca ccgacgtcgg ccgcaagctg atcatcgatc agaacgtttt
     1321 tatcgagggt acgctgccga tgggtgtcgt ccgcccgctg actgaagtcg agatggacca
     1381 ttaccgcgag ccgttcctga atcctgttga ccgcgagcca ctgtggcgct tcccaaacga
     1441 gctgccaatc gccggtgagc cagcgaacat cgtcgcgctg gtcgaagaat acatggactg
     1501 gctgcaccag tcccctgtcc cgaagctgct gttctggggc accccaggcg ttctgatccc
     1561 accggccgaa gccgctcgcc tggccaaaag cctgcctaac tgcaaggctg tggacatcgg
     1621 cccgggtctg aatctgctgc aagaagacaa cccggacctg atcggcagcg agatcgcgcg
     1681 ctggctgtcc acgctcgaga tttccggcgt gagcaagggc gaggagctgt tcaccggggt
     1741 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     1801 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     1861 caagctgccc gtgccctggc ccaccctcgt gaccaccctg tcctggggcg tgcagtgctt
     1921 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     1981 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     2041 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     2101 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tactttagcg acaacgtcta
     2161 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat
     2221 cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     2281 ccccgtgctg ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc
     2341 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     2401 cggcatggac gagctgtaca aggatgaatt gtaagcggcc gcgactctag atcataatca
     2461 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     2521 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     2581 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     2641 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     2701 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     2761 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     2821 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     2881 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     2941 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     3001 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     3061 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     3121 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3181 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3241 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     3301 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3361 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3421 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3481 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3541 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3601 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     3661 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3721 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3781 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3841 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3901 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3961 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4021 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4081 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4141 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     4201 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4261 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4321 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4381 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4441 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4501 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4561 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4621 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     4681 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     4741 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     4801 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     4861 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     4921 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4981 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5041 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5101 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5161 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5221 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5281 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5341 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5401 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5461 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5521 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5581 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5641 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgc
//
