LOCUS       sequence-470963-        8050 bp    DNA     circular SYN 01-MAY-2025
DEFINITION  Express SmBiT-PDE4C3-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 8050)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 8050)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 1, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: SmBiT-PDE4C3-LgBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..8050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3813..3831)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3868..3989
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3879..3898)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3933..3952
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4234..4591
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4234..4425
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4257..4277)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4442..4577
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4622..5659
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5638..5646
                     /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    5683..5731
                     /note="synthetic polyadenylation signal"
     promoter        5887..5991
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5992..6060,6061..6852)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6210..6229)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7010..7598)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7090..7109)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7714..7997
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgct caccatggtg accggctacc
     1081 ggctgttcga ggagattctc gggagctccg gtggtggcgg gagcggaggt ggaggctcga
     1141 gcggtgcgat cgccatgcaa ggtccaccag cgccggctcc ggttccagga cctggttctc
     1201 ctcgggggag cccgagaggc agccctggat tgttccggaa gcttttggtg aatcagagta
     1261 tcagactcca gcggaggttc accgttgctc acccgctttg cttcgatctc gaaaacggat
     1321 tgtcatgtgg aaggcgggct ctggatccac aatcaagccc ggggctcggc cggataatgc
     1381 aagcccctgt accccattct cagcgcagag agtctttctt gtaccggtct gacagtgact
     1441 atgagctgag tccgaaggcg atgtcaagaa atagtagtgt tgcgtctgac cttcacgggg
     1501 aagacatgat agtcacccct ttcgcacagg tactggcttc acttcggacg gttcgaagta
     1561 atgtagctgc actcgcacgg caacaatgtc tcggggcggc taagcaaggg cctgttggta
     1621 atcctagctc atccaatcag ctgccacctg cagaggatac aggtcagaaa ctggcacttg
     1681 aaacgttgga tgaactggac tggtgtctgg atcaactgga gacgttgcaa actagacatt
     1741 ccgtcggtga gatggcgagt aacaaattca aacggatatt gaacagagaa ctgacgcacc
     1801 tctccgaaac atctcgatcc gggaaccagg tcagcgaata tattagcaga acctttctgg
     1861 accaacaaac tgaggttgaa ctcccgaaag taactgcgga ggaggcgcca cagcccatgt
     1921 ctaggatctc cgggcttcat ggtctgtgcc actctgctag tctttcatca gcgaccgtac
     1981 cgcgatttgg ggtccaaacc gatcaagaag agcaactggc taaagaattg gaggacacaa
     2041 ataagtgggg tcttgacgtc tttaaggtcg ctgaactgag cggaaatcgc ccacttacgg
     2101 caattatctt ttctattttt caagagagag atctccttaa aacgtttcaa attccagccg
     2161 acacccttgc tacctatctc cttatgctgg agggccacta ccatgctaac gttgcttacc
     2221 ataattcact tcacgcggcg gacgtggctc aaagcacgca tgtactcctt gcaacaccgg
     2281 ccttggaagc tgtatttacg gatctggaaa tacttgctgc tttgtttgca agtgcgatcc
     2341 atgatgttga ccacccggga gtatccaacc agttcctcat taacaccaat tccgaattgg
     2401 ccctgatgta taatgacgcg tctgtgctgg aaaatcatca ccttgccgta gggttcaagt
     2461 tgctgcaggc ggagaattgc gatatcttcc agaacctttc cgccaaacag agacttagtc
     2521 tgcgccgcat ggtcattgac atggtactgg ccacggacat gagtaagcat atgaaccttt
     2581 tggccgactt gaagactatg gttgaaacga aaaaagttac aagtcttggc gtcctcttgc
     2641 tggacaatta tagcgacagg atacaggtgc tccaaaatct tgtccattgc gccgatcttt
     2701 ctaatcctac gaaacctctc ccactttacc gacagtggac ggatcggatc atggcagagt
     2761 tcttccagca aggggataga gaaagagagt ctggcttgga tatctcccca atgtgcgata
     2821 aacataccgc ctccgtggag aaaagccaag ttgggtttat agactacatt gcacacccac
     2881 tctgggaaac gtgggcagat ttggtacacc cggacgctca agaccttctg gacacactcg
     2941 aggacaatag ggagtggtat caaagcaaaa tcccccgctc acctagcgat ctgactaatc
     3001 ctgaaagaga tggtcctgac cgcttccaat tcgagctgac gctggaggaa gctgaggaag
     3061 aggatgagga ggaggaagaa gaaggggagg aaacggcact tgcgaaagag gcgctcgagc
     3121 tcccggatac agagcttctt agtcctgagg ccggaccaga tccaggggac cttcctctgg
     3181 ataatcagag aactgtttct caaggcagtt caggtggtgg cgggagcgga ggtggaggct
     3241 cgagcggtgt cttcacactc gaagatttcg ttggggactg ggaacagaca gccgcctaca
     3301 acctggacca agtccttgaa cagggaggtg tgtccagttt gctgcagaat ctcgccgtgt
     3361 ccgtaactcc gatccaaagg attgtccgga gcggtgaaaa tgccctgaag atcgacatcc
     3421 atgtcatcat cccgtatgaa ggtctgagcg ccgaccaaat ggcccagatc gaagaggtgt
     3481 ttaaggtggt gtaccctgtg gatgatcatc actttaaggt gatcctgccc tatggcacac
     3541 tggtaatcga cggggttacg ccgaacatgc tgaactattt cggacggccg tatgaaggca
     3601 tcgccgtgtt cgacggcaaa aagatcactg taacagggac cctgtggaac ggcaacaaaa
     3661 ttatcgacga gcgcctgatc acccccgacg gctccatgct gttccgagta accatcaaca
     3721 gctaaggccg cgactctaga gtcgacctgc aggcatgcaa gctgatccgg ctgctaacaa
     3781 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     3841 tggggcggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     3901 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     3961 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     4021 aggttcaggg ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta
     4081 aaatcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     4141 cgcatctgtg cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat
     4201 aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa
     4261 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4321 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4381 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4441 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4501 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4561 aggctttttt ggaggcctag gcttttgcaa aaagctcgat tcttctgaca ctagcgccac
     4621 catgaagaag cccgaactca ccgctaccag cgttgaaaaa tttctcatcg agaagttcga
     4681 cagtgtgagc gacctgatgc agttgtcgga gggcgaagag agccgagcct tcagcttcga
     4741 tgtcggcgga cgcggctatg tactgcgggt gaatagctgc gctgatggct tctacaaaga
     4801 ccgctacgtg taccgccact tcgccagcgc tgcactaccc atccccgaag tgttggacat
     4861 cggcgagttc agcgagagcc tgacatactg catcagtaga cgcgcccaag gcgttactct
     4921 ccaagacctc cccgaaacag agctgcctgc tgtgttacag cctgtcgccg aagctatgga
     4981 tgctattgcc gccgccgacc tcagtcaaac cagcggcttc ggcccattcg ggccccaagg
     5041 catcggccag tacacaacct ggcgggattt catttgcgcc attgctgatc cccatgtcta
     5101 ccactggcag accgtgatgg acgacaccgt gtccgccagc gtagctcaag ccctggacga
     5161 actgatgctg tgggccgaag actgtcccga ggtgcgccac ctcgtccatg ccgacttcgg
     5221 cagcaacaac gtcctgaccg acaacggccg catcaccgcc gtaatcgact ggtccgaagc
     5281 tatgttcggg gacagtcagt acgaggtggc caacatcttc ttctggcggc cctggctggc
     5341 ttgcatggag cagcagactc gctacttcga gcgccggcat cccgagctgg ccggcagccc
     5401 tcgtctgcga gcctacatgc tgcgcatcgg cctggatcag ctctaccaga gcctcgtgga
     5461 cggcaacttc gacgatgctg cctgggctca aggccgctgc gatgccatcg tccgcagcgg
     5521 ggccggcacc gtcggtcgca cacaaatcgc tcgccggagc gcagccgtat ggaccgacgg
     5581 ctgcgtcgag gtgctggccg acagcggcaa ccgccggccc agtacacgac cgcgcgctaa
     5641 ggaggtaggt cgagtttaga ctctagccat cacgatggcc gcaataaaat atctttattt
     5701 tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag cgataaggat cctctttgcg
     5761 cttgcgtttt cccttgtcca gatagcccag tagctgacat tcatccgggg tcagcaccgt
     5821 ttctgcggac tggctttcta cgtaatggtt tcttagacgt caggtggcac ttttcgggga
     5881 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5941 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     6001 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     6061 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     6121 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     6181 tttccaatga tgagcacttt caaagttctg ctatgtggcg cggtattatc ccgtattgac
     6241 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     6301 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6361 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaactat cggaggaccg
     6421 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6481 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6541 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6601 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6661 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6721 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6781 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6841 aagcattggt aattcgaaat gaccgaccaa gcgacgccca accggtatca gctcactcaa
     6901 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6961 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     7021 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     7081 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     7141 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     7201 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     7261 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     7321 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     7381 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     7441 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     7501 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     7561 gcaagcagca gattacgcgc agaaaaaaag gatttcaaga agatcctttg atcttttcta
     7621 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7681 caaaaaggat cttcacctag atccttttat agtccggaaa tacaggaacg cacgctggat
     7741 ggcccttcgc tgggatggtg aaaccatgaa aaatggcagc ttcagtggat taagtggggg
     7801 taatgtggcc tgtaccctct ggttgcatag gtattcatac ggttaaaatt tatcaggcgc
     7861 gattgcggca gtttttcggg tggtttgttg ccatttttac ctgtctgctg ccgtgatcgc
     7921 gctgaacgcg ttttagcggt gcgtacaatt aagggattat ggtaaatcca cttactgtct
     7981 gccctcgtag ccatcgagat aaaccgcagt actccggcca cgatgcgtcc ggcgtagagg
     8041 atcgagatct
//
