LOCUS       sequence-471846-        6350 bp    DNA     circular SYN 09-MAY-2025
DEFINITION  Express LgBiT-ARRB1 in Mammalian Cells under a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6350)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6350)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 9, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: LgBiT-ARRB1 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6350
                     /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(2230..2248)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2231..2248)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        2368..2671
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2826..2846
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2879..2926
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2958..3752
                     /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(3012..3031)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3622..3641
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      3969..4514
                     /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     4415..4434
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4631..4648
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    4798..4846
                     /note="synthetic polyadenylation signal"
     CDS             complement(4870..5907)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4883..4891
                     /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(5938..6295)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5952..6087
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6104..6295
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6252..6272
                     /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 ttaaactctg ttgttgagct
      421 gtggagagcc ggtaccatcc tcctcttcct ccttgtcatc cttcatgcct ttcagtctct
      481 ggcgagcaaa gtcctcaaat acaatgtcgt catcatttgt gtcaagttct atgagattgg
      541 tatctactgg cgtctcgttc tctggaactt cccgatgcgg gggttcctct ttgggcttgg
      601 ggtgcattag ggtgaagggc agttccacgg ccacgtcgct ggatgcaaga tctcccaaca
      661 ggccgccccg agacaccacc agcttcactt tcactttgta ggaaacaatg atccccagga
      721 tctcacggtt ggcaccttcc ctcaacaggg tgctagaggc caagttcgtg tcttcgtgct
      781 tgagcttccc gtccaaggcg aggccccgct tctctcggtt attggctagg aagggggtca
      841 gtgtgtagac cttgcagaac gtcgagctgg gtgccacagt gtcatcagcc tcttccatgg
      901 caacagggca cttgtactga gctgtgttga aaaggcagat gtctgcatac tggcgcactg
      961 agatcttgat cttcttcacc gtcttgttgg tgttgttggt gacgtggacg ttgacgctga
     1021 tgggttctcc atggtaatag atctccttat ccagagaggc ttctaggtgc aagggcttgt
     1081 ccgacatgag gaactgcctg gtggtctcgg ctgtgggctg ggggccaggc ctctctgggg
     1141 catactgaac cttccggatg accagacgca cagaattccg cttgtggatc ttctcctcca
     1201 aattctccgc gcagaaggct ttgacttcat agtccacacc gcaagccttc cccgtgtctt
     1261 cgggccccgg ctgcagtgtc acagaacatg gaaggtttgg agggatctca aaggtgaaag
     1321 ggtaagcgtg ctcgcccagc ttcttgatga ggcgttcctg cagccgcgtc aggggcttct
     1381 tgtcctcggg ggccggtggg aacgactgta cgttggccac aaacaggtcc ttgcgaaagg
     1441 tcaggcccag gacatccagg tcctcccggc catagcggaa ggcgcaggtc agcgtcacat
     1501 agactctccg ctctttgaga tactcaggat ccaccaggac cacaccatcc acagggtcca
     1561 cgaggtcgat gtggtccaca aagtcccgct ttcccaggta gacggtgagc tttccatttg
     1621 gactggcctt cttgaacact cgggtccctt tgtcgcccat ggcgatcgca ccgctcgagc
     1681 ctccacctcc gctcccgcca ccaccggagc tcccactgtt gatggttact cggaacagca
     1741 tggagccgtc gggggtgatc aggcgctcgt cgataatttt gttgccgttc cacagggtcc
     1801 ctgttacagt gatctttttg ccgtcgaaca cggcgatgcc ttcatacggc cgtccgaaat
     1861 agttcagcat gttcggcgta accccgtcga ttaccagtgt gccatagggc aggatcacct
     1921 taaagtgatg atcatccaca gggtacacca ccttaaacac ctcttcgatc tgggccattt
     1981 ggtcggcgct cagaccttca tacgggatga tgacatggat gtcgatcttc agggcatttt
     2041 caccgctccg gacaatcctt tggatcggag ttacggacac ggcgagattc tgcagcaaac
     2101 tggacacacc tccctgttca aggacttggt ccaggttgta ggcggctgtc tgttcccagt
     2161 ccccaacgaa atcttcgagt gtgaagacca tggtgagcgc tagctgcaaa aagaacaagt
     2221 agcttgtatt ctatagtgtc acctaaatgg gcgatctgac ggttcactaa accagctctg
     2281 cttatatagg cctccctggc gcgccactag ttattaatag taatcaatta cggggtcatt
     2341 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     2401 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     2461 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     2521 ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca atgacggtaa
     2581 atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta cttggcagta
     2641 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acaccaatgg
     2701 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     2761 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata accccgcccc
     2821 gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc aaatcgaatt ctaatactgt
     2881 tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta
     2941 aacccaggag gcagatcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     3001 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     3061 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     3121 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     3181 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     3241 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     3301 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     3361 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     3421 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     3481 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     3541 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     3601 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     3661 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     3721 ccttctatcg ccttcttgac gagttcttct gattcgaaat gaccgaccaa gcgacgccca
     3781 accggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3841 aggaaagaac atgtcaaaca tgagaattac aacttatatc gtatggggct gacttcaggt
     3901 gctacatttg aagagataaa ttgcactgaa atctagaaat attttatctg attaataaga
     3961 tgatcttctt gagatcgttt tggtctgcgc gtaatctctt gctctgaaaa cgaaaaaacc
     4021 gccttgcagg gcggtttttc gaaggttctc tgagctacca actctttgaa ccgaggtaac
     4081 tggcttggag gagcgcagtc accaaaactt gtcctttcag tttagcctta accggcgcat
     4141 gacttcaaga ctaactcctc taaatcaatt accagtggct gctgccagtg gtgcttttgc
     4201 atgtctttcc gggttggact caagacgata gttaccggat aaggcgcagc ggtcggactg
     4261 aacggggggt tcgtgcatac agtccagctt ggagcgaact gcctacccgg aactgagtgt
     4321 caggcgtgga atgagacaaa cgcggccata acagcggaat gacaccggta aaccgaaagg
     4381 caggaacagg agagcgcacg agggagccgc cagggggaaa cgcctggtat ctttatagtc
     4441 ctgtcgggtt tcgccaccac tgatttgagc gtcagatttc gtgatgcttg tcaggggggc
     4501 ggagcctatg gaaaaacggc tttgccgcgg ccctctcact tccctgttaa gtatcttcct
     4561 ggcatcttcc aggaaatctc cgccccgttc gtaagccatt tccgctcgcc gcagtcgaac
     4621 gaccgagcgt agcgagtcag tgagcgagga agcggaatat atcctgtatc acatattctg
     4681 ctgacgcacc ggtgcagcct tttttctcct gccacatgaa gcacttcact gacaccctca
     4741 tcagtgccaa catagtaagc cagtatacac tccgctagcg ctgatgtccg gaatcgtcac
     4801 acaaaaaacc aacacacaga tgtaatgaaa ataaagatat tttattgcgg ccatcgtgat
     4861 ggctagagtc taaactcgac ctacctcctt agcgcgcggt cgtgtactgg gccggcggtt
     4921 gccgctgtcg gccagcacct cgacgcagcc gtcggtccat acggctgcgc tccggcgagc
     4981 gatttgtgtg cgaccgacgg tgccggcccc gctgcggacg atggcatcgc agcggccttg
     5041 agcccaggca gcatcgtcga agttgccgtc cacgaggctc tggtagagct gatccaggcc
     5101 gatgcgcagc atgtaggctc gcagacgagg gctgccggcc agctcgggat gccggcgctc
     5161 gaagtagcga gtctgctgct ccatgcaagc cagccagggc cgccagaaga agatgttggc
     5221 cacctcgtac tgactgtccc cgaacatagc ttcggaccag tcgattacgg cggtgatgcg
     5281 gccgttgtcg gtcaggacgt tgttgctgcc gaagtcggca tggacgaggt ggcgcacctc
     5341 gggacagtct tcggcccaca gcatcagttc gtccagggct tgagctacgc tggcggacac
     5401 ggtgtcgtcc atcacggtct gccagtggta gacatgggga tcagcaatgg cgcaaatgaa
     5461 atcccgccag gttgtgtact ggccgatgcc ttggggcccg aatgggccga agccgctggt
     5521 ttgactgagg tcggcggcgg caatagcatc catagcttcg gcgacaggct gtaacacagc
     5581 aggcagctct gtttcgggga ggtcttggag agtaacgcct tgggcgcgtc tactgatgca
     5641 gtatgtcagg ctctcgctga actcgccgat gtccaacact tcggggatgg gtagtgcagc
     5701 gctggcgaag tggcggtaca cgtagcggtc tttgtagaag ccatcagcgc agctattcac
     5761 ccgcagtaca tagccgcgtc cgccgacatc gaagctgaag gctcggctct cttcgccctc
     5821 cgacaactgc atcaggtcgc tcacactgtc gaacttctcg atgagaaatt tttcaacgct
     5881 ggtagcggtg agttcgggct tcttcatggt ggcgctagtg tcagaagaat cgagcttttt
     5941 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga
     6001 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
     6061 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
     6121 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     6181 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     6241 ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc ctcagaaggt
     6301 acctaaccaa gttcctcttt cagaggttat ttcaggccat ggtgctgcgc
//
