LOCUS       sequence-474692-        6883 bp    DNA     circular SYN 05-JUN-2025
DEFINITION  Express TEAD2-NanoLuc(R) 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 6883)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6883)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: TEAD2-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6883
                     /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"
     CDS             complement(275..787)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     promoter        complement(2188..2206)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2189..2206)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        2326..2629
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2784..2804
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2837..2884
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2916..3575
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2982..3001)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(3747..4335)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3827..3846)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4461..4509
                     /note="synthetic polyadenylation signal"
     CDS             complement(4533..5327)
                     /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(4644..4663)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5254..5273
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5705..6883)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5714..6652
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(5737..5757)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
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 ctagagtcgc ggccttacgc cagaatgcgt tcgcacagcc
      301 gccagccggt cactccgttg atggttactc ggaacagcag ggagccgtcg gggttgatca
      361 ggcgctcgtc gataattttg ttgccgttcc acagggtccc tgttacagtg atctttttgc
      421 cgtcgaacac ggcgatgcct tcatacggcc gtccgaaata gtcgatcatg ttcggcgtaa
      481 ccccgtcgat taccagtgtg ccatagtgca ggatcacctt aaagtgatga tcatccacag
      541 ggtacaccac cttaaaaatt ttttcgatct ggcccatttg gtcgccgctc agaccttcat
      601 acgggatgat gacatggatg tcgatcttca gcccattttc accgctcagg acaatccttt
      661 ggatcggagt tacggacacc ccgagattct gaaacaaact ggacacacct ccctgttcaa
      721 ggacttggtc caggttgtag ccggctgtct gtcgccagtc cccaacgaaa tcttcgagtg
      781 tgaagacgcc gctcgagccg agagaaacgt ccctgaccag gcggtaaatg tgatgctggg
      841 ccccacgctc gctggtggag acctcgaaga cataggcggt gcagagcagc agttcctggg
      901 tgtctctgtt tgtcaccacc tggaggatgg tgaagttttc caggacgctg ttcatcatgt
      961 atcgctcagg cagctgccgc aacttgtgca agaaattcac caggtactcg cacatgggcg
     1021 agcgcagcag gcggtacaca aatctgccgt cctccagctg ggcccgttcc gtctccacct
     1081 tctccaccac ctgcttgcca aaagagcaga ccttggagga acaggtgagg gtcatgtgtt
     1141 ccaggctctc atactggctg ctcactccgt agaagccacc actgctgatg ctgccaccgg
     1201 cccctgcctc ctcaccactt gggccccagt tcaggtccgc ccagaacttg accaggaaga
     1261 aggcatgggg ggggccacga tcatatagct ctcggaggcc accctttttc tcagggaatt
     1321 tgtcgtagat ctgccggacg tccacactct cgagcggcgg cgctccgggg ctggggcagt
     1381 gctggctgat gtgcacgaac aggtgcctct ggtaagaatc aactgcatct ggcggttcca
     1441 cgaaggctga gaactctacc agctgcaacc gggcggtgcc caggccccga gcctgccagg
     1501 ctgggggcga tggggtaggt gggggcaggg gtgagagggc ttgggggggc tcgtaccctg
     1561 ggaggtcagt agatggggga gtcagtgaca aggtgaacgg tgtctgtgag aatggcttca
     1621 catctggaac attccagggg ggcccagatc ctccagacca aaactggaaa agctcagagg
     1681 cctgaggacc agtgggaccc agtttggcct gcagagaagg cgcggagatg agctgggcag
     1741 aggacatggt tgccattgtc tggaaagcct tgtccttgga aacctggtcc ttcaacttgg
     1801 actggatttc ccttgatttc cttcgggcca aaacctggat gtgactagaa acctgttttc
     1861 gagttcgggt cttccccgtt ctcagcttga tgtagcgggc gatcagttca ttccgaccat
     1921 acatcttgcc ttcatcagac aaaattattt tccggcggcc gcagggtgga tagatggcca
     1981 gggcctcctg gaagctctgc tcaatgtctg ggctccacac cccctctgca tccgggcccc
     2041 cgtcaccccc agccccctca ctgccgccgg taccctcctc actgccttcc tcactgcccg
     2101 tccagccgct gccatcgtcc agggcggccc cagcccgggg ttcccccatg gcgatcgcta
     2161 gctgcaaaaa gaacaagtag cttgtattct atagtgtcac ctaaatgggc gatctgacgg
     2221 ttcactaaac cagctctgct tatataggcc tccctggcgc gccactagtt attaatagta
     2281 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     2341 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     2401 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     2461 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagtc cgccccctat
     2521 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttacggga
     2581 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     2641 ttggcagtac accaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     2701 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     2761 tcgtaataac cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt gggaggtcaa
     2821 atcgaattct aatactgttg acaattaatc atcggcatag tatatcggca tagtataata
     2881 cgacaaggtg aggaactaaa cccaggaggc agatcatgga gaaaaaaatc actggatata
     2941 ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg
     3001 ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa
     3061 agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg
     3121 ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc
     3181 acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat
     3241 accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg
     3301 aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc
     3361 cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc
     3421 ccgttttcac tatgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga
     3481 ttcaggttca tcatgccgtc tgtgatggct tccatgtcgg cagaatgctt aatgaattac
     3541 aacagtactg cgatgagtgg cagggcgggg cgtaagcggg actctggggt tcgaaatgac
     3601 cgaccaagcg acgcccaacc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3661 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3721 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3781 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3841 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3901 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3961 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4021 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4081 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4141 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     4201 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4261 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4321 aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4381 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4441 cttttatagt ccggaatcgt cacacaaaaa accaacacac agatgtaatg aaaataaaga
     4501 tattttattg cggccatcgt gatggctaga gttcagaaga actcgtcaag aaggcgatag
     4561 aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc
     4621 cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg
     4681 tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg
     4741 atattcggca agcaggcatc gccgtgggtc acgacgagat cctcgccgtc gggcatgcgc
     4801 gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca
     4861 tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct
     4921 tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc
     4981 atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact
     5041 tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa
     5101 ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg
     5161 gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac
     5221 acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc
     5281 acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatggt ggcatcgatt
     5341 cccggtcaac atccagtaca cctccggcac accggcttcc ccaagggcgc cacgctctcc
     5401 caccacaaca tcctcaacaa cggctactgg gtgggccgca cggtcggcta cacggagcag
     5461 gaccgggtgt gtctgccggt gcccttctac cactgcttcg gcatggtcat gggcaacttg
     5521 ggcgccacct cccacgccgc ctgcatcgtc atcccggccc cgtccttcga gccggcggcg
     5581 acgctggagg cggtgcagcg ggagaggtgc acgtccctgt acggcgtccc cacgaagccg
     5641 aattccacca cactggacta gatgcatgct cgacagtgtg atggatatct gcagaattcg
     5701 gctttcacga cacctgaaat ggaagaaaaa aactttgaac cactgtctga ggcttgagaa
     5761 tgaaccaaga tccaaactca aaaagggcaa attccaagga gaattacatc aagtgccaag
     5821 ctggcctaac ttcagtctcc acccactcag tgtggggaaa ctccatcgca taaaacccct
     5881 ccccccaacc taaagacgac gtactccaaa agctcgagaa ctaatcgagg tgcctggacg
     5941 gcgcccggta ctccgtggag tcacatgaag cgacggctga ggacggaaag gcccttttcc
     6001 tttgtgtggg tgactcaccc gcccgctctc ccgagcgccg cgtcctccat tttgagctgc
     6061 ctgcagcagg gccgggaagc ggccatcttt ccgctcacgc aactggtgcc gaccgggcca
     6121 gccttgccgc ccagggcggg gcgatacacg gcggcgcgag gccaggcacc agagcaggcc
     6181 ggccagcttg agactacccc cgtccgattc tcggtggccg cgctcgcagg ccccgcctcg
     6241 ccgaacatgt gcgctgggac gcacgggccc cgtcgccgcc cgcggcccca aaaaccgaaa
     6301 taccagtgtg cagatcttgg cccgcattta caagactatc ttgccagaaa aaaagcgtcg
     6361 cagcaggtca tcaaaaattt taaatggcta gagacttatc gaaagcagcg agacaggcgc
     6421 gaaggtgcca ccagattcgc acgcggcggc cccagcgccc aggccaggcc tcaactcaag
     6481 cacgaggcga aggggctcct taagcgcaag gcctcgaact ctcccaccca cttccaaccc
     6541 gaagctcggg atcaagaatc acgtactgca gccaggtgga agtaattcaa ggcacgcaag
     6601 ggccataacc cgtaaagagg ccaggcccgc gggaaccaca cacggcactt acctgtgttc
     6661 tggcggcaaa cccgttgcga aaaagaacgt tcacggcgac tactgcactt atatacggtt
     6721 ctcccccacc ctcgggaaaa aggcggagcc agtacacgac atcactttcc cagtttaccc
     6781 cgcgccacct tctctaggca ccggttcaat tgccgacccc tccccccaac ttctcgggga
     6841 ctgtgggcga tgtgcgctct gcccactgac gggcaccgga gcc
//
