LOCUS       sequence-481492-        6820 bp    DNA     circular SYN 30-JUL-2025
DEFINITION  Express TEAD1-NanoLuc 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 6820)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6820)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 30, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: TEAD1-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6820
                     /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(2125..2143)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2126..2143)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        2263..2566
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2721..2741
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2774..2821
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2853..3512
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2919..2938)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(3684..4272)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3764..3783)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4398..4446
                     /note="synthetic polyadenylation signal"
     CDS             complement(4470..5264)
                     /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(4581..4600)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5191..5210
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5642..6820)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5651..6589
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(5674..5694)
                     /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 cctttacaag cctgtaaata tgatgttgtg
      841 ctccgtgttc actatttgaa acttcaaaca cacaggccat gcagagtaga gtttcttgtg
      901 tatccctgtt tgttaccacc aataaaattg tgaagttttc caaaacactg ttcatcatat
      961 atttctctgg taagtgtttg agcttgtgga tgaagttgat catatattca cacattgggg
     1021 agcggtttat tcggtataca aatcggccat tctcaaacct tgcatactcc gtctctactt
     1081 tttctactac ttgcttccca aaggagcaaa ctttggtgga acaggtgact gtcatatttt
     1141 cagaactctc gtactgactg gttacaccat aaaaagcccc agcatcatct tgaatattgc
     1201 agtttaaatc agcccagaat tttacgagga agaaggcatt ttgagggccc tttccaaaca
     1261 gttcctttaa gccacctttc ttttcaggaa atttgtcata aatctgacga atgtccactg
     1321 attcaagcaa tgggtcactg taagaatggt tggcatgccc aatgtgcacg aagaggtgtt
     1381 tgttgtacga gtctgggtct cgctgctgct cgagaaaagc tgaaaattcc accaggcgaa
     1441 gcttggttgt gccaatggag cgaccttgcc aggcagggac tgagggagct ggggccgatg
     1501 caggctcaaa ccctggaatg ggggctgtga ccgctggctg gatggggtag gcctgctgca
     1561 caaaaggctt gacgtcttgt gaggatcctg gctgccctgt ttgaatcatt cccggccaga
     1621 accccggcgc ccctgggaag gtcgggcgtg gaatcccagg cagccccagc ttgttatgaa
     1681 tggcagtggc cgagacgatc tgggctgagg acatggccgc catgtgctgc agggccttat
     1741 cctttgcagt ctgatccttt agcttggaat gaaaatcacg agatttcctt ctggcaagaa
     1801 cctgaatgtg actagacacc tgttttctgg tcctcgtctt gcctgtcctg agtttgatgt
     1861 atctggctat caattcattc ctaccataca ttttgccttc gtctgataag atgattttcc
     1921 tcctcccaca tggtggatag atagccaggg cctcctgaaa gctttgctcg atgtcggggc
     1981 tccagacccc ttctgcatca ttgtcaattg gcttatctgc agagtcactc atcctttcca
     2041 tgttttcggc agggctctca ctgccgctcc agctgctggg ctccatggcg atcgctagct
     2101 gcaaaaagaa caagtagctt gtattctata gtgtcaccta aatgggcgat ctgacggttc
     2161 actaaaccag ctctgcttat ataggcctcc ctggcgcgcc actagttatt aatagtaatc
     2221 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     2281 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     2341 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     2401 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtccgc cccctattga
     2461 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tacgggactt
     2521 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     2581 gcagtacacc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     2641 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     2701 taataacccc gccccgttga cgcaaatggg cggtaggcgt gtacggtggg aggtcaaatc
     2761 gaattctaat actgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
     2821 caaggtgagg aactaaaccc aggaggcaga tcatggagaa aaaaatcact ggatatacca
     2881 ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc
     2941 aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga
     3001 aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc
     3061 atccggaatt ccgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc
     3121 cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc
     3181 acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa
     3241 acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct
     3301 gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg
     3361 ttttcactat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc
     3421 aggttcatca tgccgtctgt gatggcttcc atgtcggcag aatgcttaat gaattacaac
     3481 agtactgcga tgagtggcag ggcggggcgt aagcgggact ctggggttcg aaatgaccga
     3541 ccaagcgacg cccaaccggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3601 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3661 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3721 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3781 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3841 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3901 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3961 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4021 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4081 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     4141 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4201 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4261 aaaggatttc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4321 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4381 ttatagtccg gaatcgtcac acaaaaaacc aacacacaga tgtaatgaaa ataaagatat
     4441 tttattgcgg ccatcgtgat ggctagagtt cagaagaact cgtcaagaag gcgatagaag
     4501 gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat
     4561 tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc
     4621 gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata
     4681 ttcggcaagc aggcatcgcc gtgggtcacg acgagatcct cgccgtcggg catgcgcgcc
     4741 ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc
     4801 tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg
     4861 tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg
     4921 atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg
     4981 cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga
     5041 acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cctgcagttc attcagggca
     5101 ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg
     5161 gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc
     5221 caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatggtggc atcgattccc
     5281 ggtcaacatc cagtacacct ccggcacacc ggcttcccca agggcgccac gctctcccac
     5341 cacaacatcc tcaacaacgg ctactgggtg ggccgcacgg tcggctacac ggagcaggac
     5401 cgggtgtgtc tgccggtgcc cttctaccac tgcttcggca tggtcatggg caacttgggc
     5461 gccacctccc acgccgcctg catcgtcatc ccggccccgt ccttcgagcc ggcggcgacg
     5521 ctggaggcgg tgcagcggga gaggtgcacg tccctgtacg gcgtccccac gaagccgaat
     5581 tccaccacac tggactagat gcatgctcga cagtgtgatg gatatctgca gaattcggct
     5641 ttcacgacac ctgaaatgga agaaaaaaac tttgaaccac tgtctgaggc ttgagaatga
     5701 accaagatcc aaactcaaaa agggcaaatt ccaaggagaa ttacatcaag tgccaagctg
     5761 gcctaacttc agtctccacc cactcagtgt ggggaaactc catcgcataa aacccctccc
     5821 cccaacctaa agacgacgta ctccaaaagc tcgagaacta atcgaggtgc ctggacggcg
     5881 cccggtactc cgtggagtca catgaagcga cggctgagga cggaaaggcc cttttccttt
     5941 gtgtgggtga ctcacccgcc cgctctcccg agcgccgcgt cctccatttt gagctgcctg
     6001 cagcagggcc gggaagcggc catctttccg ctcacgcaac tggtgccgac cgggccagcc
     6061 ttgccgccca gggcggggcg atacacggcg gcgcgaggcc aggcaccaga gcaggccggc
     6121 cagcttgaga ctacccccgt ccgattctcg gtggccgcgc tcgcaggccc cgcctcgccg
     6181 aacatgtgcg ctgggacgca cgggccccgt cgccgcccgc ggccccaaaa accgaaatac
     6241 cagtgtgcag atcttggccc gcatttacaa gactatcttg ccagaaaaaa agcgtcgcag
     6301 caggtcatca aaaattttaa atggctagag acttatcgaa agcagcgaga caggcgcgaa
     6361 ggtgccacca gattcgcacg cggcggcccc agcgcccagg ccaggcctca actcaagcac
     6421 gaggcgaagg ggctccttaa gcgcaaggcc tcgaactctc ccacccactt ccaacccgaa
     6481 gctcgggatc aagaatcacg tactgcagcc aggtggaagt aattcaaggc acgcaagggc
     6541 cataacccgt aaagaggcca ggcccgcggg aaccacacac ggcacttacc tgtgttctgg
     6601 cggcaaaccc gttgcgaaaa agaacgttca cggcgactac tgcacttata tacggttctc
     6661 ccccaccctc gggaaaaagg cggagccagt acacgacatc actttcccag tttaccccgc
     6721 gccaccttct ctaggcaccg gttcaattgc cgacccctcc ccccaacttc tcggggactg
     6781 tgggcgatgt gcgctctgcc cactgacggg caccggagcc
//
