LOCUS       sequence-474951-        7098 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express SLC35A2-HiBiT in Mammalian Cells under a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7098)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7098)
  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            "Unpublished"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: SLC35A2-HiBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..7098
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        66..257
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     214..234
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    336..417
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        650..953
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        954..1157
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1108..1128
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          1293..1425
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1408..1427
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1469..1488
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1469..1487
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3466..3486
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             3502..4386
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     polyA_signal    4473..4594
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4484..4503)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4709..4742)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4752..4799
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4831..5625
                     /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(4885..4904)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5495..5514
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5783..6371)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5863..5882)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6497..6545
                     /note="synthetic polyadenylation signal"
     CDS             complement(6569..6967)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6964..6973
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(join(6998..7098,1..257))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(7012..7098,1..49)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta
       61 ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
      121 gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac
      181 ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagct
      241 ggttctttcc gcctcagaag gtacctaacc aagttcctct ttcagaggtt atttcaggcc
      301 atggtgctgc gcaagacgcg ttaattaaga cgtcggcagt gaaaaaaatg ctttatttgt
      361 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
      421 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
      481 agcaagtaaa acctctacaa atgtggtatg gctgattatg atccgcggcc gcaatacgtc
      541 gacgttatca gctgacttcg tacgagagcc taggattatg gcgcgccact agttattaat
      601 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
      661 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      721 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      781 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc
      841 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac
      901 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
      961 ggttttggca gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     1021 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     1081 aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg
     1141 tctatataag cagagctggt ttagtgaacc gtcagatcac tagaagcttt attgcggtag
     1201 tttatcacag ttaaattgct aacgcagtca gtgcttctga cacaacagtc tcgaacttaa
     1261 gctgcagaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca agacaggttt
     1321 aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg
     1381 cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagt
     1441 tcaattacag ctcttaaggc tagagtatta atacgactca ctatagggct agcgatcgcc
     1501 atggagcaca tacagggagc ttggaagacg atcagcaatg gttttggatt caaagatgcc
     1561 gtgtttgatg gctccagctg catctctcct acaatagttc agcagtttgg ctatcagcgc
     1621 cgggcatcag atgatggcaa actcacagat ccttctaaga caagcaacac tatccgtgtt
     1681 ttcttgccga acaagcaaag aacagtggtc aatgtgcgaa atggaatgag cttgcatgac
     1741 tgccttatga aagcactcaa ggtgaggggc ctgcaaccag agtgctgtgc agtgttcaga
     1801 cttctccacg aacacaaagg taaaaaagca cgcttagatt ggaatactga tgctgcgtct
     1861 ttgattggag aagaacttca agtagatttc ctggatcatg ttcccctcac aacacacaac
     1921 tttgctcgga agacgttcct gaagcttgcc ttctgtgaca tctgtcagaa attcctgctc
     1981 aatggatttc gatgtcagac ttgtggctac aaatttcatg agcactgtag caccaaagta
     2041 cctactatgt gtgtggactg gagtaacatc agacaactct tattgtttcc aaattccact
     2101 attggtgata gtggagtccc agcactacct tctttgacta tgcgtcgtat gcgagagtct
     2161 gtttccagga tgcctgttag ttctcagcac agatattcta cacctcacgc cttcaccttt
     2221 aacacctcca gtccctcatc tgaaggttcc ctctcccaga ggcagaggtc gacatccaca
     2281 cctaatgtcc acatggtcag caccaccctg cctgtggaca gcaggatgat tgaggatgca
     2341 attcgaagtc acagcgaatc agcctcacct tcagccctgt ccagtagccc caacaatctg
     2401 agcccaacag gctggtcaca gccgaaaacc cccgtgccag cacaaagaga gcgggcacca
     2461 gtatctggga cccaggagaa aaacaaaatt aggcctcgtg gacagagaga ttcaagctat
     2521 tattgggaaa tagaagccag tgaagtgatg ctgtccactc ggattgggtc aggctctttt
     2581 ggaactgttt ataagggtaa atggcacgga gatgttgcag taaagatcct aaaggttgtc
     2641 gacccaaccc cagagcaatt ccaggccttc aggaatgagg tggctgttct gcgcaaaaca
     2701 cggcatgtga acattctgct tttcatgggg tacatgacaa aggacaacct ggcaattgtg
     2761 acccagtggt gcgagggcag cagcctctac aaacacctgc atgtccagga gaccaagttt
     2821 cagatgttcc agctaattga cattgcccgg cagacggctc agggaatgga ctatttgcat
     2881 gcaaagaaca tcatccatag agacatgaaa tccaacaata tatttctcca tgaaggctta
     2941 acagtgaaaa ttggagattt tggtttggca acagtaaagt cacgctggag tggttctcag
     3001 caggttgaac aacctactgg ctctgtcctc tggatggccc cagaggtgat ccgaatgcag
     3061 gataacaacc cattcagttt ccagtcggat gtctactcct atggcatcgt attgtatgaa
     3121 ctgatgacgg gggagcttcc ttattctcac atcaacaacc gagatcagat catcttcatg
     3181 gtgggccgag gatatgcctc cccagatctt agtaagctat ataagaactg ccccaaagca
     3241 atgaagaggc tggtagctga ctgtgtgaag aaagtaaagg aagagaggcc tctttttccc
     3301 cagatcctgt cttccattga gctgctccaa cactctctac cgaagatcaa ccggagcgct
     3361 tccgagccat ccttgcatcg ggcagcccac actgaggata tcaatgcttg cacgctgacc
     3421 acgtccccga ggctgcctgt cttcgtttct ctcgagccaa ccactgagga tctgtacttt
     3481 cagagcgata acgatggatc cgaaatcggt actggctttc cattcgaccc ccattatgtg
     3541 gaagtcctgg gcgagcgcat gcactacgtc gatgttggtc cgcgcgatgg cacccctgtg
     3601 ctgttcctgc acggtaaccc gacctcctcc tacgtgtggc gcaacatcat cccgcatgtt
     3661 gcaccgaccc atcgctgcat tgctccagac ctgatcggta tgggcaaatc cgacaaacca
     3721 gacctgggtt atttcttcga cgaccacgtc cgcttcatgg atgccttcat cgaagccctg
     3781 ggtctggaag aggtcgtcct ggtcattcac gactggggct ccgctctggg tttccactgg
     3841 gccaagcgca atccagagcg cgtcaaaggt attgcattta tggagttcat ccgccctatc
     3901 ccgacctggg acgaatggcc agaatttgcc cgcgagacct tccaggcctt ccgcaccacc
     3961 gacgtcggcc gcaagctgat catcgatcag aacgttttta tcgagggtac gctgccgatg
     4021 ggtgtcgtcc gcccgctgac tgaagtcgag atggaccatt accgcgagcc gttcctgaat
     4081 cctgttgacc gcgagccact gtggcgcttc ccaaacgagc tgccaatcgc cggtgagcca
     4141 gcgaacatcg tcgcgctggt cgaagaatac atggactggc tgcaccagtc ccctgtcccg
     4201 aagctgctgt tctggggcac cccaggcgtt ctgatcccac cggccgaagc cgctcgcctg
     4261 gccaaaagcc tgcctaactg caaggctgtg gacatcggcc cgggtctgaa tctgctgcaa
     4321 gaagacaacc cggacctgat cggcagcgag atcgcgcgct ggctgtctac tctggagatt
     4381 tccggttaat agaattctag agtcgacctg caggcatgca agctgatttg gctgctgcca
     4441 ccgctgagca aggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     4501 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     4561 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     4621 gtttcaggtt cagggggaga tgtgggaggt ttttttaagc aagtaaaacc tctacaaatg
     4681 tggtaaaatc ttaattaaga attctaatat aacttcgtat agcatacatt atacgaagtt
     4741 atggatccac tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
     4801 aggtgaggaa ctaaacccag gaggcagatc atgattgaac aagatggatt gcacgcaggt
     4861 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4921 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4981 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     5041 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     5101 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     5161 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     5221 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5281 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     5341 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5401 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5461 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5521 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5581 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgattcga aatgaccgac
     5641 caagcgacgc ccaaccggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5701 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5761 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5821 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5881 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5941 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6001 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6061 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6121 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6181 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     6241 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6301 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6361 aaggatttca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6421 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6481 tatagtccgg aatcgtcaca caaaaaacca acacacagat gtaatgaaaa taaagatatt
     6541 ttattgcggc catcgtgatg gctagagttt agccctccca cacataacca gagggcagca
     6601 attcacgaat cccaactgcc gtcggctgtc catcactgtc cttcactatg gctttgatcc
     6661 caggatgcag atcgagaagc acctgtcggc accgtccgca ggggctcaag atgcccctgt
     6721 tctcatttcc gatcgcgacg atacaagtca ggttgccagc tgccgcagca gcagcagtgc
     6781 ccagcaccac gagttctgca caaggtcccc cagtaaaatg atatacattg acaccagtga
     6841 agatgcggcc gtcgctagag agagctgcgc tggcgacgct gtagtcttca gagatgggga
     6901 tgctgttgat tgtagccgtt gctctttcaa tgagggtgga ttcttcttga gacaaaggct
     6961 tggccatggt ggcgctagtg tcagaagaat cgagcttttt gcaaaagcct aggcctccaa
     7021 aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat
     7081 aaataaaaaa aattagtc
//
