LOCUS       sequence-474590-        5594 bp    DNA     circular SYN 04-JUN-2025
DEFINITION  Express LgBiT-PDE4C3-SmBiT 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 5594)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5594)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 4, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: LgBiT-PDE4C3-SmBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5594
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        87..390
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        510..528
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     510..527
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1883..1901)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2078..2159
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2115..2134)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2231..2588
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2231..2422
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2254..2274)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2439..2574
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2619..3656
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3635..3643
                     /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)"
     polyA_signal    3680..3728
                     /note="synthetic polyadenylation signal"
     rep_origin      3854..4442
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4343..4362
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4600..5394)
                     /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(4711..4730)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5321..5340
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5426..5473)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     primer_bind     complement(5506..5526)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca
       61 ttggtgtact gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac
      121 tgccaagtag gaaagtcccg taaggtcatg tactgggcat aatgccaggc gggccattta
      181 ccgtcattga cgtcaatagg gggcggactt ggcatatgat acacttgatg tactgccaag
      241 tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt
      301 actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca
      361 ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat
      421 gaccccgtaa ttgattacta ttaataacta gtggcgcgcc agggaggcct atataagcag
      481 agctggttta gtgaaccgtc agatcgccca tttaggtgac actatagaat acaagctact
      541 tgttcttttt gcagctagcg ctcaccatgg tcttcacact cgaagatttc gttggggact
      601 gggaacagac agccgcctac aacctggacc aagtccttga acagggaggt gtgtccagtt
      661 tgctgcagaa tctcgccgtg tccgtaactc cgatccaaag gattgtccgg agcggtgaaa
      721 atgccctgaa gatcgacatc catgtcatca tcccgtatga aggtctgagc gccgaccaaa
      781 tggcccagat cgaagaggtg tttaaggtgg tgtaccctgt ggatgatcat cactttaagg
      841 tgatcctgcc ctatggcaca ctggtaatcg acggggttac gccgaacatg ctgaactatt
      901 tcggacggcc gtatgaaggc atcgccgtgt tcgacggcaa aaagatcact gtaacaggga
      961 ccctgtggaa cggcaacaaa attatcgacg agcgcctgat cacccccgac ggctccatgc
     1021 tgttccgagt aaccatcaac agtgggagct ccggtggtgg cgggagcgga ggtggaggct
     1081 cgagcggtgc gatcgccatg tcggtgttcg ggaagctgtt cggggctgga gggggtaagg
     1141 ccggcaaggg cggcccgacc ccccaggagg ccatccagcg gctgcgggac acggaagaga
     1201 tgttaagcaa gaaacaggag ttcctggaga agaaaatcga gcaggagctg acggccgcca
     1261 agaagcacgg caccaaaaac aagcgcgcgg ccctccaggc actgaagcgt aagaagaggt
     1321 atgagaagca gctggcgcag atcgacggca cattatcaac catcgagttc cagcgggagg
     1381 ccctggagaa tgccaacacc aacaccgagg tgctcaagaa catgggctat gccgccaagg
     1441 ccatgaaggc ggcccatgac aacatggaca tcgataaagt tgatgagtta atgcaggaca
     1501 ttgctgacca gcaagaactt gcagaggaga tttcaacagc aatttcgaaa cctgtagggt
     1561 ttggagaaga gtttgacgag gatgagctca tggcggaatt agaagaacta gaacaggagg
     1621 aactagacaa gaatttgctg gaaatcagtg gacccgaaac agtccctcta ccaaatgttc
     1681 cctctatagc cctaccatca aaacccgcca agaagaaaga agaggaggac gacgacatga
     1741 aggaattgga gaactgggct ggatccatgg tttaaacgaa ttcgggctcg gtacccgggg
     1801 atcctctaga gtcgacctgc aggcatgcaa gctgatccgg ctgctaacaa agcccgaaag
     1861 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggaaaca
     1921 cctaggctct cgtacgaagt cagctgataa cgtcgacgta ttgcggccgc ggatcataat
     1981 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     2041 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     2101 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgcc
     2161 gacgtcacgc gtcttgcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt
     2221 taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     2281 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2341 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2401 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2461 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2521 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     2581 tttgcaaaaa gctcgattct tctgacacta gcgccaccat gaagaagccc gaactcaccg
     2641 ctaccagcgt tgaaaaattt ctcatcgaga agttcgacag tgtgagcgac ctgatgcagt
     2701 tgtcggaggg cgaagagagc cgagccttca gcttcgatgt cggcggacgc ggctatgtac
     2761 tgcgggtgaa tagctgcgct gatggcttct acaaagaccg ctacgtgtac cgccacttcg
     2821 ccagcgctgc actacccatc cccgaagtgt tggacatcgg cgagttcagc gagagcctga
     2881 catactgcat cagtagacgc gcccaaggcg ttactctcca agacctcccc gaaacagagc
     2941 tgcctgctgt gttacagcct gtcgccgaag ctatggatgc tattgccgcc gccgacctca
     3001 gtcaaaccag cggcttcggc ccattcgggc cccaaggcat cggccagtac acaacctggc
     3061 gggatttcat ttgcgccatt gctgatcccc atgtctacca ctggcagacc gtgatggacg
     3121 acaccgtgtc cgccagcgta gctcaagccc tggacgaact gatgctgtgg gccgaagact
     3181 gtcccgaggt gcgccacctc gtccatgccg acttcggcag caacaacgtc ctgaccgaca
     3241 acggccgcat caccgccgta atcgactggt ccgaagctat gttcggggac agtcagtacg
     3301 aggtggccaa catcttcttc tggcggccct ggctggcttg catggagcag cagactcgct
     3361 acttcgagcg ccggcatccc gagctggccg gcagccctcg tctgcgagcc tacatgctgc
     3421 gcatcggcct ggatcagctc taccagagcc tcgtggacgg caacttcgac gatgctgcct
     3481 gggctcaagg ccgctgcgat gccatcgtcc gcagcggggc cggcaccgtc ggtcgcacac
     3541 aaatcgctcg ccggagcgca gccgtatgga ccgacggctg cgtcgaggtg ctggccgaca
     3601 gcggcaaccg ccggcccagt acacgaccgc gcgctaagga ggtaggtcga gtttagactc
     3661 tagccatcac gatggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt
     3721 tttgtgtgac gattccggac tataaaagga tctaggtgaa gatccttttt gataatctca
     3781 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3841 tcaaaggatc ttcttgaaat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3901 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3961 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     4021 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4081 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4141 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4201 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4261 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4321 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4381 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4441 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4501 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     4561 ctgataccgg ttgggcgtcg cttggtcggt catttcgaat cagaagaact cgtcaagaag
     4621 gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg
     4681 gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg
     4741 atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc
     4801 caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg
     4861 catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc
     4921 cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg
     4981 tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc
     5041 atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc
     5101 cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc
     5161 tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cctgcagttc
     5221 attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag
     5281 ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag
     5341 cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgatctg
     5401 cctcctgggt ttagttcctc accttgtcgt attatactat gccgatatac tatgccgatg
     5461 attaattgtc aacagtatta gaattcgatt tgacctccca ccgtacacgc ctaccgccca
     5521 tttgcgtcaa cggggcgggg ttattacgac attttggaaa gtcccgttga ttttggtgcc
     5581 aaaacaaact ccca
//
