LOCUS       sequence-473315-        5377 bp    DNA     circular SYN 18-JUN-2025
DEFINITION  Express LgBiT-CYBC1 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 5377)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5377)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 18, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: LgBiT-CYBC1 BiBiT-Ready Vector (CMV)
FEATURES             Location/Qualifiers
     source          1..5377
                     /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(1431..1449)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1432..1449)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        1569..1872
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2027..2047
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2080..2127
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2159..2953
                     /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(2213..2232)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2823..2842
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3111..3699)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3191..3210)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3825..3873
                     /note="synthetic polyadenylation signal"
     CDS             complement(3897..4934)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3910..3918
                     /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(4965..5322)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4979..5114
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5131..5322
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5279..5299
                     /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 ctagaggatc cccgggtacc gagcccgaat tcgtttaaac
      301 gctctggctt gcagggtcac cggcctcact gtcgctgctc tgagacagct ctgtggggct
      361 ctcaaggcag tgcagctcca ggaagctggt gatgagcttg gcgatggctt ccacatcact
      421 gcggtggccc atgactgcac tctgcgtgag ggggtgggag aagcccgtcg caagccggag
      481 caccaccatg tagcctttcc cgaagtaccg gaccttctcc tcctccacgc tcacatcacg
      541 gacatcatgg agcaggacca ccacctggtc gtggccagct ctgaaaagag tcagcagctt
      601 cttgtagagg ctgaacgtct tcaaaacaac cttccctgtg ctcttgtcga agatggcttc
      661 ctcccagtcc tccaagttct gcacagccac aaacaggcag cctgtgacgt agaagagctt
      721 ccagcccagg ctatctccgc tgtagtaggc agcagccagg ccaatcgaca agattccaac
      781 cagcagggac caggaccgga tgcctggagc cctcttcaga tggaggcggg agctggtgcg
      841 ggtctccacc tgcaggtaca tggcgatcgc accgctcgag cctccacctc cgctcccgcc
      901 accaccggag ctcccactgt tgatggttac tcggaacagc atggagccgt cgggggtgat
      961 caggcgctcg tcgataattt tgttgccgtt ccacagggtc cctgttacag tgatcttttt
     1021 gccgtcgaac acggcgatgc cttcatacgg ccgtccgaaa tagttcagca tgttcggcgt
     1081 aaccccgtcg attaccagtg tgccataggg caggatcacc ttaaagtgat gatcatccac
     1141 agggtacacc accttaaaca cctcttcgat ctgggccatt tggtcggcgc tcagaccttc
     1201 atacgggatg atgacatgga tgtcgatctt cagggcattt tcaccgctcc ggacaatcct
     1261 ttggatcgga gttacggaca cggcgagatt ctgcagcaaa ctggacacac ctccctgttc
     1321 aaggacttgg tccaggttgt aggcggctgt ctgttcccag tccccaacga aatcttcgag
     1381 tgtgaagacc atggtgagcg ctagctgcaa aaagaacaag tagcttgtat tctatagtgt
     1441 cacctaaatg ggcgatctga cggttcacta aaccagctct gcttatatag gcctccctgg
     1501 cgcgccacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     1561 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     1621 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     1681 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     1741 catatgccaa gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     1801 gcccagtaca tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc
     1861 gctattacca tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac
     1921 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     1981 aatcaacggg actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt
     2041 aggcgtgtac ggtgggaggt caaatcgaat tctaatactg ttgacaatta atcatcggca
     2101 tagtatatcg gcatagtata atacgacaag gtgaggaact aaacccagga ggcagatcat
     2161 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     2221 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     2281 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     2341 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     2401 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     2461 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     2521 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     2581 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     2641 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     2701 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     2761 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     2821 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     2881 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     2941 cgagttcttc tgattcgaaa tgaccgacca agcgacgccc aaccggtatc agctcactca
     3001 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3061 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3121 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3181 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3241 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3301 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3361 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3421 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3481 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3541 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3601 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3661 tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag aagatccttt gatcttttct
     3721 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3781 tcaaaaagga tcttcaccta gatcctttta tagtccggaa tcgtcacaca aaaaaccaac
     3841 acacagatgt aatgaaaata aagatatttt attgcggcca tcgtgatggc tagagtctaa
     3901 actcgaccta cctccttagc gcgcggtcgt gtactgggcc ggcggttgcc gctgtcggcc
     3961 agcacctcga cgcagccgtc ggtccatacg gctgcgctcc ggcgagcgat ttgtgtgcga
     4021 ccgacggtgc cggccccgct gcggacgatg gcatcgcagc ggccttgagc ccaggcagca
     4081 tcgtcgaagt tgccgtccac gaggctctgg tagagctgat ccaggccgat gcgcagcatg
     4141 taggctcgca gacgagggct gccggccagc tcgggatgcc ggcgctcgaa gtagcgagtc
     4201 tgctgctcca tgcaagccag ccagggccgc cagaagaaga tgttggccac ctcgtactga
     4261 ctgtccccga acatagcttc ggaccagtcg attacggcgg tgatgcggcc gttgtcggtc
     4321 aggacgttgt tgctgccgaa gtcggcatgg acgaggtggc gcacctcggg acagtcttcg
     4381 gcccacagca tcagttcgtc cagggcttga gctacgctgg cggacacggt gtcgtccatc
     4441 acggtctgcc agtggtagac atggggatca gcaatggcgc aaatgaaatc ccgccaggtt
     4501 gtgtactggc cgatgccttg gggcccgaat gggccgaagc cgctggtttg actgaggtcg
     4561 gcggcggcaa tagcatccat agcttcggcg acaggctgta acacagcagg cagctctgtt
     4621 tcggggaggt cttggagagt aacgccttgg gcgcgtctac tgatgcagta tgtcaggctc
     4681 tcgctgaact cgccgatgtc caacacttcg gggatgggta gtgcagcgct ggcgaagtgg
     4741 cggtacacgt agcggtcttt gtagaagcca tcagcgcagc tattcacccg cagtacatag
     4801 ccgcgtccgc cgacatcgaa gctgaaggct cggctctctt cgccctccga caactgcatc
     4861 aggtcgctca cactgtcgaa cttctcgatg agaaattttt caacgctggt agcggtgagt
     4921 tcgggcttct tcatggtggc gctagtgtca gaagaatcga gctttttgca aaagcctagg
     4981 cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc
     5041 tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga
     5101 gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc
     5161 atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac
     5221 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     5281 cctaactgac acacattcca cagctggttc tttccgcctc agaaggtacc taaccaagtt
     5341 cctctttcag aggttatttc aggccatggt gctgcgc
//
