LOCUS       sequence-472422-        8236 bp    DNA     circular SYN 13-MAY-2025
DEFINITION  Express AR-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 8236)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 8236)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: CMV-AR-SmBiT Expression Vector
FEATURES             Location/Qualifiers
     source          1..8236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(579..936)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      593..728
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        745..936
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     893..913
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    1181..1302
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1218..1237)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1272..1291
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1339..1357
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     primer_bind     complement(4304..4323)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4305..4323)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4365..4384)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          4367..4499
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(4635..4838)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4664..4684)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4839..5218
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    5409..5692
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     rep_origin      5808..6396
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6297..6316
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6554..7345,7346..7414))
                     /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     7177..7196
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7415..7519)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    7675..7723
                     /note="synthetic polyadenylation signal"
     CDS             complement(join(7747..8236,1..548))
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             7760..7768
                     /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)"
ORIGIN
        1 agcatcagtt cgtccagggc ttgagctacg ctggcggaca cggtgtcgtc catcacggtc
       61 tgccagtggt agacatgggg atcagcaatg gcgcaaatga aatcccgcca ggttgtgtac
      121 tggccgatgc cttggggccc gaatgggccg aagccgctgg tttgactgag gtcggcggcg
      181 gcaatagcat ccatagcttc ggcgacaggc tgtaacacag caggcagctc tgtttcgggg
      241 aggtcttgga gagtaacgcc ttgggcgcgt ctactgatgc agtatgtcag gctctcgctg
      301 aactcgccga tgtccaacac ttcggggatg ggtagtgcag cgctggcgaa gtggcggtac
      361 acgtagcggt ctttgtagaa gccatcagcg cagctattca cccgcagtac atagccgcgt
      421 ccgccgacat cgaagctgaa ggctcggctc tcttcgccct ccgacaactg catcaggtcg
      481 ctcacactgt cgaacttctc gatgagaaat ttttcaacgc tggtagcggt gagttcgggc
      541 ttcttcatgg tggcgctagt gtcagaagaa tcgagctttt tgcaaaagcc taggcctcca
      601 aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca
      661 taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg
      721 ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt
      781 tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg
      841 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac
      901 tgacacacat tccacagctg gttctttccg cctcagaagg tacctaacca agttcctctt
      961 tcagaggtta tttcaggcca tggtgctgcg cagatccgcg tatgcggtgt gaaataccgc
     1021 acagatgcgt aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa
     1081 attcgatttt accacatttg tagaggtttt acttgcttaa aaaaacctcc cacatctccc
     1141 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     1201 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     1261 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gctcgaagcg
     1321 gccgccccaa ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc
     1381 tttcgggctt tgttagcagc cggatcagct tgcatgcctg caggtcgact ctagagtcgc
     1441 ggccttacag aatctcctcg aacagccggt agccggtcac accgctcgag cctccacctc
     1501 cgctcccgcc accacctgaa ctgccttgag aaacctgggt gtggaaatag atgggcttga
     1561 ctttcccaga aaggatcttg ggcacttgca cagagatgat ctctgccatc atttccggaa
     1621 agtccacgct caccatgtgt gacttgatta gcaggtcaaa agtgaactga tgcagctctc
     1681 tcgcaatagg ctgcacggag tccaggagct tggtgagctg gtagaagcgt cttgagcagg
     1741 atgtgggatt ttttcttttg catgcaatga tacgatcgag ttccttgatg tagttcattc
     1801 gaagttcatc aaagaatttt tgatttttca gcccatccac tggaataatg ctgaagagta
     1861 gcagtgcttt catgcacagg aattcctggg gggtgatttg gagccatcca aactcttgag
     1921 agaggtgcct cattcggaca cactggctgt acatccggga cttgtgcatg cggtactcat
     1981 tgaaaaccag atcaggggcg aagtagagca tcctggagtt gacattggtg aaggatcgcc
     2041 agcccatggc aaacaccatg agccccatcc aggagtactg aatgacagcc atctggtcgt
     2101 ccacgtgtaa gttgcggaag ccaggcaagg ccttggccca cttgaccacg tgtacaagct
     2161 gtctctctcc cagttcattg aggctagaga gcaaggctgc aaaggagtcg ggctggttgt
     2221 tgtcgtgtcc agcacacact acacctggct caatggcttc caggacattc agaaagatgg
     2281 gctgacattc atagccttca atgtgtgaca ctgtcagctt ctgggttgtc tcctcagtgg
     2341 ggctggtggt gctggaagcc tctccttcct cctgtagttt cagattacca agtttcttca
     2401 gcttccgggc tcccagagtc atccctgctt cataacattt ccgaagacga caagatggac
     2461 aatttttcct tcggaattta tcaatagtgc aatcatttct gctggcgcac aggtacttct
     2521 gtttcccttc agcggctctt ttgaagaaga ccttgcagct tccacatgtg agagctccat
     2581 agtgacaccc agaagcttca tctccacaga tcaggcaggt cttctggggt ggaaagtaat
     2641 agtcaatggg caaaacatgg tccctggcag tctccaaacg catgtccccg taaggtccgg
     2701 agtagctatc catccagggg cccatttcgc ttttgacaca agtgggactg ggatagggca
     2761 ctctgctcac catgccgcca gggtaccaca catcaggtgc ggtgaagtcg ctttcctggc
     2821 ccgccagccc ctgagggggc cgagtgtagc cgtagggggc tacagctccc gcctcgccgc
     2881 cgccgccgcc gccgccgccg ccgccgccgc cgccgccgcc gccgccgcca ccacccccac
     2941 caccaccaca cggtccatac aactggcctt cttcggctgt gaagagagtg tgccaggatg
     3001 aggaagcggc ggctgagggt gacccagaac cgggtcccgc tgcacccgcg ccatgcaggc
     3061 tcgccaggtc cccatagcgg cactgcgccg ccgcagccgc ccaggcgctg ccgtagtcca
     3121 gcgggttctc cagcttgatg cgagcgtggg gatggggagg cggcggaggg ggcggcggtc
     3181 cggccagagc cagtggaaag ttgtagtagt cgcgactctg gtacgcagct gcctcgtcca
     3241 gtgctccgga cttgtagaga gacagggtag acggcagttc aagtgtcccg gagctccctg
     3301 ctgcagcgct gccagagcag cctaggctct cgccttctag ccctttggtg taacctccct
     3361 tgaaagggga atactcagca gtatcttcag tgctcttgcc tgcgctgtcg tctagcagag
     3421 aacctttgca ttcggccaat ggggcacaag gagtgggacg cacagcgggt ggaactccca
     3481 aaagtggggc gtacatgcaa tccccccgaa gctgttcccc tggactcaga tgctccaacg
     3541 cctccacacc caggcccatg gacaccgaca ctgccttaca caactccttg gcgttgtcag
     3601 aaatggtcga agtgccccct aagtaattgt ccttggagga agtgggagcc cccgaggctt
     3661 ccctcgctct cccgctgctg ctgccttcgg atactgcttc ctgctgctgt tgctgaagga
     3721 gttgcatggt gctggcctcg ctcaggatgt ctttaaggtc agcggagcag ctgcttaagc
     3781 cggggaaagt ggggcccagc agggacaacg tggatggggc agctgagtca tcctcgtccg
     3841 gaggtgctgg cagctgctgc ggcagcccct tgctggcggc cacggcggct ccaggctctg
     3901 ggacgcaacc tctctcgggg tggcactcca gggccgactg cggctgtgaa ggttgctgtt
     3961 cctcatccag gaccaggtag cctgtggggc ctctacgatg ggcttgggga gaaccatcct
     4021 caccctgctg ctgctgctgc tgcctggggc tagtctcttg ctgctgctgc tgctgctgct
     4081 gctgctgctg ctgctgctgc tgctgctgct gctgctgctg cagcagcagc aaactggcgc
     4141 cgggaggtgc tgcgctcgcg gcctctgggt gcctggggcc cgggttctgg atcacttcgc
     4201 gcacgctctg gaacagattc tggaaagctc ctcggtaggt cttggacggc ggccgagggt
     4261 agacccttcc cagccctaac tgcacttcca tggcgatcgc tagccctata gtgagtcgta
     4321 ttaatactct agccttaaga gctgtaattg aactgggagt ggacacctgt ggagagaaag
     4381 gcaaagtgga tgtcagtaag accaataggt gcctatcaga aacgcaagag tcttctctgt
     4441 ctcgacaagc ccagtttcta ttggtctcct taaacctgtc ttgtaacctt gatacttacc
     4501 tgcccagtgc ctcacgacca acttctgcag cttaagttcg agactgttgt gtcagaagca
     4561 ctgactgcgt tagcaattta actgtgataa actaccgcaa taaagcttct agtgatctga
     4621 cggttcacta aaccagctct gcttatatag acctcccacc gtacacgcct accgcccatt
     4681 tgcgtcaacg gggcggggtt attacgacat tttggaaagt cccgttgatt ttggtgccaa
     4741 aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc
     4801 tatccacgcc cattggtgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat
     4861 acgtagatgt actgccaagt aggaaagtcc cgtaaggtca tgtactgggc ataatgccag
     4921 gcgggccatt taccgtcatt gacgtcaata gggggcggac ttggcatatg atacacttga
     4981 tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc
     5041 cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg
     5101 ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg
     5161 ctatgaacta atgaccccgt aattgattac tattaataac tagtcaataa tcaatgccaa
     5221 catggcggtc atattggaca tgagccaata taaatgtaca tattatgata tagatacaac
     5281 gtatgcaatg gccaatagcc aatattgatt tatgctatat aaccaatgaa taatatggct
     5341 aatggccaat attgaagatc tcgatcctct acgccggacg catcgtggcc ggagtactgc
     5401 ggtttatctc gatggctacg agggcagaca gtaagtggat ttaccataat cccttaattg
     5461 tacgcaccgc taaaacgcgt tcagcgcgat cacggcagca gacaggtaaa aatggcaaca
     5521 aaccacccga aaaactgccg caatcgcgcc tgataaattt taaccgtatg aatacctatg
     5581 caaccagagg gtacaggcca cattaccccc acttaatcca ctgaagctgc catttttcat
     5641 ggtttcacca tcccagcgaa gggccatcca gcgtgcgttc ctgtatttcc ggactataaa
     5701 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5761 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg aaatcctttt
     5821 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5881 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5941 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6001 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6061 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6121 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6181 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6241 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6301 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6361 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6421 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6481 tctgtggata accgtattac cgcctttgag tgagctgata ccggttgggc gtcgcttggt
     6541 cggtcatttc gaattaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6601 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6661 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6721 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6781 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6841 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6901 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6961 tgtgcaaaaa agcggttagc tccttcggtc ctccgatagt tgtcagaagt aagttggccg
     7021 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7081 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7141 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7201 ctttgaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7261 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7321 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7381 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7441 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7501 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     7561 ttacgtagaa agccagtccg cagaaacggt gctgaccccg gatgaatgtc agctactggg
     7621 ctatctggac aagggaaaac gcaagcgcaa agaggatcct tatcgctatc gattcacaca
     7681 aaaaaccaac acacagatgt aatgaaaata aagatatttt attgcggcca tcgtgatggc
     7741 tagagtctaa actcgaccta cctccttagc gcgcggtcgt gtactgggcc ggcggttgcc
     7801 gctgtcggcc agcacctcga cgcagccgtc ggtccatacg gctgcgctcc ggcgagcgat
     7861 ttgtgtgcga ccgacggtgc cggccccgct gcggacgatg gcatcgcagc ggccttgagc
     7921 ccaggcagca tcgtcgaagt tgccgtccac gaggctctgg tagagctgat ccaggccgat
     7981 gcgcagcatg taggctcgca gacgagggct gccggccagc tcgggatgcc ggcgctcgaa
     8041 gtagcgagtc tgctgctcca tgcaagccag ccagggccgc cagaagaaga tgttggccac
     8101 ctcgtactga ctgtccccga acatagcttc ggaccagtcg attacggcgg tgatgcggcc
     8161 gttgtcggtc aggacgttgt tgctgccgaa gtcggcatgg acgaggtggc gcacctcggg
     8221 acagtcttcg gcccac
//
