LOCUS       sequence-471896-        6284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express LgBiT-SLC35A2 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 6284)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6284)
  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            "Unpublished"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: LgBiT-SLC35A2 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    17..98
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     42..61
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     275..293
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     promoter        complement(2164..2182)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2165..2182)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        2302..2605
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2760..2780
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2813..2860
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2892..3686
                     /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(2946..2965)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3556..3575
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      3903..4448
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     4349..4368
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4565..4582
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    4732..4780
                     /note="synthetic polyadenylation signal"
     CDS             complement(4804..5841)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4817..4825
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(5872..6229)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5886..6021
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6038..6229
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6186..6206
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtg acgtcggcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
       61 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      121 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
      181 aatgtggtat ggctgattat gatccgcggc cgcaatacgt cgacgttatc agctgacttc
      241 gtacgagagc ctaggtgttt ccccaagggg ttatgctagt tattgctcag cggtggcagc
      301 agccaactca gcttcctttc gggctttgtt agcagccgga tcagcttgca tgcctgcagg
      361 tcgactctag aggatccccg ggtaccgagc ccgaattcgt ttaaacggat cccttcactt
      421 tggtcagcaa tttcggcagg aagggctcgg ttatgaggtc accgcggtgg gaggacaact
      481 gtggaggcgg cggctgtccc ggaggttgct gatggacgca cggaccggag gcagacgcgg
      541 aagcagaggc tatggctttc gctgctcccc gaggcaggga gtacaaatag accgctccta
      601 tgacaagccc agcacccaag gcaaagagcg gatctacatg aaacccgaag agtcttattg
      661 aggcgacggt ggacaatacg atggacaagg aggtcgcgaa tcccttcagt atgttgtccg
      721 cgtatttcac aacaaccgcc acgagcaatc cgccaaatgc ttggttcaat acgacacccc
      781 aaacagctgg tgtgtatcca aaaaagaatc cgcgagttgc gactgcggtg ccctccgccc
      841 accacaaacc caccaacccg agggccgtcc caaagaggcc aagctgcaga tttctcaacc
      901 agacactacc agaagaaccc ttcaggatct tctcaaaata tacacccgcg aacccagagc
      961 taaggcagga ggccaccaca gccgcgagcc cggctcccgg attctgatcg agagggcggg
     1021 gacctcctcc tcccgcttgt tgagcttgaa ctatggcgac ccctgtaaag aggagaagga
     1081 gactggccca ttggagtcgg gacaaggacc ggttcagcat aagaacggag aacaatgccg
     1141 ttgtcaggat tttcagttgg taggtgactt ggaacgtagc tgccggcaaa tttgatatag
     1201 cgacatattg aagattattt tgcaaagtat aaatgaggct gggtaccgcc aacttcagtg
     1261 tgtcaacata ttggacaagc actgcctcgt gcagaaacag aaccaggtgt tttacatttc
     1321 ctcgtttttg cgcaaagaga aggagcaggc acgtcaatcc cttaaggacc tctgccatga
     1381 cgactgcggt tgttgcgaaa aatcgatcgc ccggcaacgt ccgagcgtat cttatagaaa
     1441 ggatgaggga agcattttgc actacgagaa cagcaagaga aatatacttc aatcttcggt
     1501 gggcggctga agcagtccca ggttcaagag cgcctgcaga caccgcgccc ggaccaggtg
     1561 ctgccgtact tcctcctgcg cccacggcag ccatggcgat cgcaccgctc gagcctccac
     1621 ctccgctccc gccaccaccg gagctcccac tgttgatggt tactcggaac agcatggagc
     1681 cgtcgggggt gatcaggcgc tcgtcgataa ttttgttgcc gttccacagg gtccctgtta
     1741 cagtgatctt tttgccgtcg aacacggcga tgccttcata cggccgtccg aaatagttca
     1801 gcatgttcgg cgtaaccccg tcgattacca gtgtgccata gggcaggatc accttaaagt
     1861 gatgatcatc cacagggtac accaccttaa acacctcttc gatctgggcc atttggtcgg
     1921 cgctcagacc ttcatacggg atgatgacat ggatgtcgat cttcagggca ttttcaccgc
     1981 tccggacaat cctttggatc ggagttacgg acacggcgag attctgcagc aaactggaca
     2041 cacctccctg ttcaaggact tggtccaggt tgtaggcggc tgtctgttcc cagtccccaa
     2101 cgaaatcttc gagtgtgaag accatggtga gcgctagctg caaaaagaac aagtagcttg
     2161 tattctatag tgtcacctaa atgggcgatc tgacggttca ctaaaccagc tctgcttata
     2221 taggcctccc tggcgcgcca ctagttatta atagtaatca attacggggt cattagttca
     2281 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     2341 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     2401 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     2461 acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc
     2521 cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta
     2581 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg
     2641 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     2701 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac
     2761 gcaaatgggc ggtaggcgtg tacggtggga ggtcaaatcg aattctaata ctgttgacaa
     2821 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccca
     2881 ggaggcagat catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     2941 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3001 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3061 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3121 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     3181 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     3241 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3301 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3361 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     3421 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     3481 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     3541 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     3601 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     3661 atcgccttct tgacgagttc ttctgattcg aaatgaccga ccaagcgacg cccaaccggt
     3721 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3781 gaacatgtca aacatgagaa ttacaactta tatcgtatgg ggctgacttc aggtgctaca
     3841 tttgaagaga taaattgcac tgaaatctag aaatatttta tctgattaat aagatgatct
     3901 tcttgagatc gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg
     3961 cagggcggtt tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt
     4021 ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc
     4081 aagactaact cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct
     4141 ttccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg
     4201 gggttcgtgc atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg
     4261 tggaatgaga caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa
     4321 caggagagcg cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg
     4381 ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc
     4441 tatggaaaaa cggctttgcc gcggccctct cacttccctg ttaagtatct tcctggcatc
     4501 ttccaggaaa tctccgcccc gttcgtaagc catttccgct cgccgcagtc gaacgaccga
     4561 gcgtagcgag tcagtgagcg aggaagcgga atatatcctg tatcacatat tctgctgacg
     4621 caccggtgca gccttttttc tcctgccaca tgaagcactt cactgacacc ctcatcagtg
     4681 ccaacatagt aagccagtat acactccgct agcgctgatg tccggaatcg tcacacaaaa
     4741 aaccaacaca cagatgtaat gaaaataaag atattttatt gcggccatcg tgatggctag
     4801 agtctaaact cgacctacct ccttagcgcg cggtcgtgta ctgggccggc ggttgccgct
     4861 gtcggccagc acctcgacgc agccgtcggt ccatacggct gcgctccggc gagcgatttg
     4921 tgtgcgaccg acggtgccgg ccccgctgcg gacgatggca tcgcagcggc cttgagccca
     4981 ggcagcatcg tcgaagttgc cgtccacgag gctctggtag agctgatcca ggccgatgcg
     5041 cagcatgtag gctcgcagac gagggctgcc ggccagctcg ggatgccggc gctcgaagta
     5101 gcgagtctgc tgctccatgc aagccagcca gggccgccag aagaagatgt tggccacctc
     5161 gtactgactg tccccgaaca tagcttcgga ccagtcgatt acggcggtga tgcggccgtt
     5221 gtcggtcagg acgttgttgc tgccgaagtc ggcatggacg aggtggcgca cctcgggaca
     5281 gtcttcggcc cacagcatca gttcgtccag ggcttgagct acgctggcgg acacggtgtc
     5341 gtccatcacg gtctgccagt ggtagacatg gggatcagca atggcgcaaa tgaaatcccg
     5401 ccaggttgtg tactggccga tgccttgggg cccgaatggg ccgaagccgc tggtttgact
     5461 gaggtcggcg gcggcaatag catccatagc ttcggcgaca ggctgtaaca cagcaggcag
     5521 ctctgtttcg gggaggtctt ggagagtaac gccttgggcg cgtctactga tgcagtatgt
     5581 caggctctcg ctgaactcgc cgatgtccaa cacttcgggg atgggtagtg cagcgctggc
     5641 gaagtggcgg tacacgtagc ggtctttgta gaagccatca gcgcagctat tcacccgcag
     5701 tacatagccg cgtccgccga catcgaagct gaaggctcgg ctctcttcgc cctccgacaa
     5761 ctgcatcagg tcgctcacac tgtcgaactt ctcgatgaga aatttttcaa cgctggtagc
     5821 ggtgagttcg ggcttcttca tggtggcgct agtgtcagaa gaatcgagct ttttgcaaaa
     5881 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
     5941 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
     6001 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
     6061 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
     6121 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     6181 tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcaga aggtacctaa
     6241 ccaagttcct ctttcagagg ttatttcagg ccatggtgct gcgc
//
