LOCUS       sequence-473317-        5345 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express TUBEs-LgBiT 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 5345)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5345)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: TUBEs-LgBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5345
                     /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"
     promoter        complement(1399..1417)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1400..1417)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        1537..1840
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1995..2015
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2048..2095
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2127..2921
                     /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(2181..2200)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2791..2810
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3079..3667)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3159..3178)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3793..3841
                     /note="synthetic polyadenylation signal"
     CDS             complement(3865..4902)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3878..3886
                     /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(4933..5290)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4947..5082
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5099..5290
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5247..5267
                     /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 ctagagtcgc ggccttagct gttgatggtt actcggaaca
      301 gcatggagcc gtcgggggtg atcaggcgct cgtcgataat tttgttgccg ttccacaggg
      361 tccctgttac agtgatcttt ttgccgtcga acacggcgat gccttcatac ggccgtccga
      421 aatagttcag catgttcggc gtaaccccgt cgattaccag tgtgccatag ggcaggatca
      481 ccttaaagtg atgatcatcc acagggtaca ccaccttaaa cacctcttcg atctgggcca
      541 tttggtcggc gctcagacct tcatacggga tgatgacatg gatgtcgatc ttcagggcat
      601 tttcaccgct ccggacaatc ctttggatcg gagttacgga cacggcgaga ttctgcagca
      661 aactggacac acctccctgt tcaaggactt ggtccaggtt gtaggcggct gtctgttccc
      721 agtccccaac gaaatcttcg agtgtgaaga ccatgccaga acctcctccc ccggatcctc
      781 ctccgccgga ccctccacca ccgcctgtca gcaaatactc gacagctcga tgaggattgt
      841 tataggaggc ccgcagggca gctacaacgc gctcgcgttc gtaacccatt gacattatct
      901 cagtaagcat agtctcgtat tcggatccgc tgccacttcc gctcccaccg gtcagcaggt
      961 attcaaccgc cctatgcgga ttattatatg aggctcgcag cgcagcgacc actctctccc
     1021 tctcgtatcc catactcata atctcggtga gcatcgtctc atattcactg ccgctacctg
     1081 atccactacc acccgtcagc agatactcaa cagctctatg ggggttatta taggaagctc
     1141 ggagcgctgc gacaactcgt tcccgttcat agcccatgga cataatctct gtcagcatgg
     1201 tctcatactc tgaacctgac ccgcttccac tccctcctgt cagaagatat tccaccgctc
     1261 ggtgaggatt gttgtagctc gcgcgcagag ccgccacaac gcgctcccgt tcatatccca
     1321 ttgacattat ttccgtgagc atggtttcat attcggacat ggcgatcgct agctgcaaaa
     1381 agaacaagta gcttgtattc tatagtgtca cctaaatggg cgatctgacg gttcactaaa
     1441 ccagctctgc ttatataggc ctccctggcg cgccactagt tattaatagt aatcaattac
     1501 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     1561 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     1621 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     1681 tgcccacttg gcagtacatc aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa
     1741 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttacggg actttcctac
     1801 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     1861 caccaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     1921 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa
     1981 ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtca aatcgaattc
     2041 taatactgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     2101 gaggaactaa acccaggagg cagatcatga ttgaacaaga tggattgcac gcaggttctc
     2161 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     2221 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     2281 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     2341 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     2401 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     2461 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     2521 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     2581 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     2641 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     2701 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     2761 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     2821 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     2881 agcgcatcgc cttctatcgc cttcttgacg agttcttctg attcgaaatg accgaccaag
     2941 cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3001 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3061 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3121 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3181 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3241 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3301 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3361 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3421 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3481 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     3541 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3601 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3661 atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3721 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttata
     3781 gtccggaatc gtcacacaaa aaaccaacac acagatgtaa tgaaaataaa gatattttat
     3841 tgcggccatc gtgatggcta gagtctaaac tcgacctacc tccttagcgc gcggtcgtgt
     3901 actgggccgg cggttgccgc tgtcggccag cacctcgacg cagccgtcgg tccatacggc
     3961 tgcgctccgg cgagcgattt gtgtgcgacc gacggtgccg gccccgctgc ggacgatggc
     4021 atcgcagcgg ccttgagccc aggcagcatc gtcgaagttg ccgtccacga ggctctggta
     4081 gagctgatcc aggccgatgc gcagcatgta ggctcgcaga cgagggctgc cggccagctc
     4141 gggatgccgg cgctcgaagt agcgagtctg ctgctccatg caagccagcc agggccgcca
     4201 gaagaagatg ttggccacct cgtactgact gtccccgaac atagcttcgg accagtcgat
     4261 tacggcggtg atgcggccgt tgtcggtcag gacgttgttg ctgccgaagt cggcatggac
     4321 gaggtggcgc acctcgggac agtcttcggc ccacagcatc agttcgtcca gggcttgagc
     4381 tacgctggcg gacacggtgt cgtccatcac ggtctgccag tggtagacat ggggatcagc
     4441 aatggcgcaa atgaaatccc gccaggttgt gtactggccg atgccttggg gcccgaatgg
     4501 gccgaagccg ctggtttgac tgaggtcggc ggcggcaata gcatccatag cttcggcgac
     4561 aggctgtaac acagcaggca gctctgtttc ggggaggtct tggagagtaa cgccttgggc
     4621 gcgtctactg atgcagtatg tcaggctctc gctgaactcg ccgatgtcca acacttcggg
     4681 gatgggtagt gcagcgctgg cgaagtggcg gtacacgtag cggtctttgt agaagccatc
     4741 agcgcagcta ttcacccgca gtacatagcc gcgtccgccg acatcgaagc tgaaggctcg
     4801 gctctcttcg ccctccgaca actgcatcag gtcgctcaca ctgtcgaact tctcgatgag
     4861 aaatttttca acgctggtag cggtgagttc gggcttcttc atggtggcgc tagtgtcaga
     4921 agaatcgagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa
     4981 tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
     5041 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
     5101 gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     5161 tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     5221 tgctggggag cctggggact ttccacaccc taactgacac acattccaca gctggttctt
     5281 tccgcctcag aaggtaccta accaagttcc tctttcagag gttatttcag gccatggtgc
     5341 tgcgc
//
