LOCUS       sequence-473359-        7683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express TTC7B-HaloTag(R) Fusion Protein in Mammalian Cells under a 
            CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7683)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7683)
  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            "Unpublished"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: TTC7B-HaloTag Fusion Vector
FEATURES             Location/Qualifiers
     source          1..7683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    61..182
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(72..91)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(297..330)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        340..387
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             419..1213
                     /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(473..492)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1083..1102
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(1371..1959)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1451..1470)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2085..2133
                     /note="synthetic polyadenylation signal"
     CDS             complement(2157..2555)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      2552..2561
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2586..2943)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2600..2735
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        2752..2943
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     2900..2920
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    3022..3103
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        3336..3639
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3640..3843
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3794..3814
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          3979..4111
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4094..4113
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     4155..4174
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4155..4173
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6737..6757
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             6773..7657
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
ORIGIN
        1 ggcatgcaag ctgatttggc tgctgccacc gctgagcaag gccgcttcga gcagacatga
       61 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
      121 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
      181 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
      241 ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatctt aattaagaat tctaatataa
      301 cttcgtatag catacattat acgaagttat ggatccactg ttgacaatta atcatcggca
      361 tagtatatcg gcatagtata atacgacaag gtgaggaact aaacccagga ggcagatcat
      421 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
      481 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
      541 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
      601 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
      661 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
      721 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
      781 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
      841 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
      901 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
      961 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     1021 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     1081 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     1141 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     1201 cgagttcttc tgattcgaaa tgaccgacca agcgacgccc aaccggtatc agctcactca
     1261 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     1321 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1381 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1441 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     1501 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     1561 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     1621 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     1681 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     1741 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     1801 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     1861 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     1921 tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag aagatccttt gatcttttct
     1981 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2041 tcaaaaagga tcttcaccta gatcctttta tagtccggaa tcgtcacaca aaaaaccaac
     2101 acacagatgt aatgaaaata aagatatttt attgcggcca tcgtgatggc tagagtttag
     2161 ccctcccaca cataaccaga gggcagcaat tcacgaatcc caactgccgt cggctgtcca
     2221 tcactgtcct tcactatggc tttgatccca ggatgcagat cgagaagcac ctgtcggcac
     2281 cgtccgcagg ggctcaagat gcccctgttc tcatttccga tcgcgacgat acaagtcagg
     2341 ttgccagctg ccgcagcagc agcagtgccc agcaccacga gttctgcaca aggtccccca
     2401 gtaaaatgat atacattgac accagtgaag atgcggccgt cgctagagag agctgcgctg
     2461 gcgacgctgt agtcttcaga gatggggatg ctgttgattg tagccgttgc tctttcaatg
     2521 agggtggatt cttcttgaga caaaggcttg gccatggtgg cgctagtgtc agaagaatcg
     2581 agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca
     2641 gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga
     2701 gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg
     2761 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
     2821 tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
     2881 gagcctgggg actttccaca ccctaactga cacacattcc acagctggtt ctttccgcct
     2941 cagaaggtac ctaaccaagt tcctctttca gaggttattt caggccatgg tgctgcgcaa
     3001 gacgcgttaa ttaagacgtc ggcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3061 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3121 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     3181 ctacaaatgt ggtatggctg attatgatcc gcggccgcaa tacgtcgacg ttatcagctg
     3241 acttcgtacg agagcctagg attatggcgc gccactagtt attaatagta atcaattacg
     3301 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     3361 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     3421 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     3481 gcccacttgg cagtacatca agtgtatcat atgccaagtc cgccccctat tgacgtcaat
     3541 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttacggga ctttcctact
     3601 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     3661 accaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     3721 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaataac
     3781 cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     3841 gctggtttag tgaaccgtca gatcactaga agctttattg cggtagttta tcacagttaa
     3901 attgctaacg cagtcagtgc ttctgacaca acagtctcga acttaagctg cagaagttgg
     3961 tcgtgaggca ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag
     4021 aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt
     4081 actgacatcc actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct
     4141 taaggctaga gtattaatac gactcactat agggctagcg atcgccatgg cgaccaagaa
     4201 ggcaggctcg cggctggaga cggagatcga gcgctgccgc tccgagtgcc agtgggagcg
     4261 gatccctgag ctcgtcaagc agctgtcggc caagctcatc gccaacgatg acatggcaga
     4321 gcttctcctc ggggagtcga agctggagca gtacctgaag gaacaccccc tgaggcaggg
     4381 ggccagtccc cgaggcccca agccccagct gactgaggtc cgcaagcatc tgaccgccgc
     4441 cctggaccga gggaacctta agtcagaatt cctacaagaa tccaatctga tcatggccaa
     4501 gttgaattat gtggaaggtg attataaaga agctctgaac atttacgccc gggtgggcct
     4561 ggacgatctg ccactgacag ctgtcccgcc ctacaggctg cgggtgatcg cagaagccta
     4621 cgctaccaaa ggactttgtt tggagaagct gcctatttct tcttctacca gtaatctcca
     4681 tgtggaccgg gaacaggatg tcatcacctg ttatgagaaa gcaggggaca tcgcactcct
     4741 gtatctccaa gagatagaaa gggtaatact ttctaatatt caaaacagaa gccctaagcc
     4801 tggccctgct ccccacgatc aagaactagg ttttttccta gaaacaggac ttcagagagc
     4861 ccatgtcctc tatttcaaaa atgggaactt gacaagagga gtcggaagat ttagagagct
     4921 tctcagagca gttgaaacaa gaacgactca aaacctgcga atgacaatag ccaggcagct
     4981 ggcagagatc ttgttgcggg gtatgtgtga gcagagctac tggaaccctc tggaggatcc
     5041 accgtgccag tcacctctgg acgatcctct ccgcaaagga gcaaacacaa aaacctacac
     5101 tctcactcgg agagcccgtg tctactcagg agagaacatt ttttgtcctc aagaaaatac
     5161 ggaagaagcc ctgttgttat tgctgattag tgaatcaatg gccaaccggg acgctgtgct
     5221 gagcaggata cctgaacaca agagtgaccg cctcatcagt ctgcagagtg catctgtggt
     5281 ctatgactta ctcaccattg ctcttggaag aagaggccag tatgagatgc tgtcagagtg
     5341 cctagaaaga gccatgaagt ttgcctttga ggaattccac ctgtggtacc agtttgctct
     5401 gtccctgatg gctgctggaa aatctgcccg tgccgtgaag gtgctgaaag agtgtatccg
     5461 cctgaagcca gacgatgcca ccatccctct cctcgctgcc aagctctgca tgggctccct
     5521 gcactggttg gaagaggctg aaaagtttgc caaaactgtc gttgatgtgg gagagaaaac
     5581 gtcagagttc aaggccaaag gctacttagc tctggggctc acgtacagtc tgcaggccac
     5641 tgacgcttct ttgcgaggga tgcaggaggt cctacagaga aaggcgcttc ttgcatttca
     5701 gagggcccac agcctgtcac ccacagatca ccaagcagct ttctacctgg ctctgcagct
     5761 tgccatctcc agacagatcc cagaggctct ggggtatgtc cgccaagctc ttcagcttca
     5821 aggtgacgat gccaactccc tgcacctcct tgccctcctg ctgtcagcac agaagcatta
     5881 ccatgacgct ctgaacatca tcgacatggc cctgagtgaa tacccagaaa atttcatact
     5941 actgttttcc aaagtgaagt tgcagtcact ctgccgaggc ccggacgagg cactgctgac
     6001 ttgtaagcac atgctgcaga tatggaaatc ctgctacaac ctcaccaacc ccagtgattc
     6061 tggacgtggg agcagcctct tagatagaac cattgctgac agacgacagc ttaatacaat
     6121 tactttgcca gacttcagcg atcccgagac aggctccgtc catgccacat cggtagcagc
     6181 ctcaagagtg gagcaggcac tgtcggaagt ggcttcgtct ctgcagagca gtgcccctaa
     6241 gcagggcccg ctgcacccct ggatgacgct ggcacagatc tggctccatg cagctgaagt
     6301 ctatatcggc atcgggaagc ctgcagaagc cacagcctgt acccaagaag ctgccaacct
     6361 cttcccaatg tcccacaatg tcctctacat gcgcggccag attgctgagc tccggggaag
     6421 catggacgag gcgcggcggt ggtatgaaga ggccttagcc atcagcccca cccacgtgaa
     6481 gagcatgcag cgactggccc tgatccttca ccagctaggc cgctacagtc tggcggagaa
     6541 gatcctccgg gacgcggtgc aggtgaactc gacagcccac gaggtctgga acgggctggg
     6601 cgaggtcctc caagctcagg gcaacgatgc ggcggctacg gagtgcttcc tgacagcctt
     6661 ggagctggag gccagcagcc ccgccgtgcc cttcaccatc atcccccgcg tgctcgtttc
     6721 tctcgagcca accactgagg atctgtactt tcagagcgat aacgatggat ccgaaatcgg
     6781 tactggcttt ccattcgacc cccattatgt ggaagtcctg ggcgagcgca tgcactacgt
     6841 cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc cgacctcctc
     6901 ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc catcgctgca ttgctccaga
     6961 cctgatcggt atgggcaaat ccgacaaacc agacctgggt tatttcttcg acgaccacgt
     7021 ccgcttcatg gatgccttca tcgaagccct gggtctggaa gaggtcgtcc tggtcattca
     7081 cgactggggc tccgctctgg gtttccactg ggccaagcgc aatccagagc gcgtcaaagg
     7141 tattgcattt atggagttca tccgccctat cccgacctgg gacgaatggc cagaatttgc
     7201 ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc cgcaagctga tcatcgatca
     7261 gaacgttttt atcgagggta cgctgccgat gggtgtcgtc cgcccgctga ctgaagtcga
     7321 gatggaccat taccgcgagc cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt
     7381 cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata
     7441 catggactgg ctgcaccagt cccctgtccc gaagctgctg ttctggggca ccccaggcgt
     7501 tctgatccca ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt
     7561 ggacatcggc ccgggtctga atctgctgca agaagacaac ccggacctga tcggcagcga
     7621 gatcgcgcgc tggctgtcta ctctggagat ttccggttaa tagaattcta gagtcgacct
     7681 gca
//
