LOCUS       sequence-470509-        8189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express E-cadherin1-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 8189)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8189)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: E-cadherin1-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..8189
                     /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(3493..3511)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3494..3511)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        3631..3934
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4089..4109
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        4142..4189
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4221..4880
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4287..4306)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(5052..5640)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5132..5151)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5766..5814
                     /note="synthetic polyadenylation signal"
     CDS             complement(5838..6632)
                     /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(5949..5968)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6559..6578
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(7011..8189)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          7020..7958
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(7043..7063)
                     /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 agagtttggt cgtcctcgcc gcctccgtac atgtcagcca
      841 gcttcttgaa gcgattgccc cattcgttca agtagtcata gtcctggtct ttgtctgact
      901 ctgaggagtt cagggagctc agactagcag cttcggaacc gcttccttca tagtcaaaca
      961 cgagcagaga atcataaggc ggggctgtgg ggtcagtatc agccgctttc agattttcat
     1021 caataaaatt tccaatttca tcgggattgg cagggcgggg aagataccgg gggacactca
     1081 tgagggttgg tgcaacgtcg ttacgagtca cttcaggccg agcgtccagg cccctgtgca
     1141 gctggctcaa gtcaaagtcc tggtcctctt ctccgcctcc ttcttcatca tagtaataaa
     1201 cgttgtcccg ggtgtcatcc tctgggggca gtaagggctc tttgaccacc gctctcctcc
     1261 gaagaaacag caagagcagc agaatcagaa ttagcaaagc aagaattcct ccaagaatcc
     1321 ccagaatggc aggaatttgc aatcctgctt cgacaggctg tgccttccta cagacgccag
     1381 cggccccttc acagtcacac acgctgacct ctaaggtggt cacttggtct ttattctggt
     1441 tatccatgag cttgagattg attttgtagt cacccacctc taaggccatc tttggcttca
     1501 aaatgataga ttcttgggtt gggtcgttgt actgaatggt ccagttggca ctcgccccgt
     1561 gtgttagttc tgctgtgaag ggagatgtat tgggaggaag gtctgcatca atgatgttta
     1621 tgacctgagg ctttggattc ctctcacaga agaatatagt tcgaggttct ggtatggggg
     1681 cgttgtcatt cacatcagac aggatcagca gaagtgtccc tgttccagta gcaactggag
     1741 aaccattgtc tgtagctatg attagggctg tgtacgtgct gttcttcacg tgctcaaaat
     1801 cctccctgtc cagctcagcc cgagtggaaa tggcaccagt gtccggatta atctccagcc
     1861 agttggcagt gtctctccaa atccgatatg ttattttctg ttccataaat gtgtctggct
     1921 cctgggcagt gtaggatgtg atttcctggc ccacgccaaa gtcctcggac acttccactc
     1981 tcttttcagg aggcacaaag atgggggctt cattcacatc cagcacatcc acggtgacgg
     2041 tggctgtgga ggtggtgaga gagacctcaa aaggtaccac attcgtcact gctacgtgta
     2101 gaatgtactg ctgcttggcc tcaaaatcca agccctttgc tgttttcaaa atgccatcgt
     2161 tgttcactgg atttgtggtg acgacaaatt gtccaccatc atcattcaat atggtgtata
     2221 cagcctccca cgctggggta ttgggggcat cagcatcagt cactttcagt gtggtgatta
     2281 cgacgttagc ctcgttctca ggcacctgac ccttgtacgt ggtgggattg aagatcggag
     2341 gattatcgtt ggtgtcagtg actgtgatca cagctgttgc tgttgtgctt aacccctcac
     2401 cttgaaggtc agcagcttga accaccaggg tatacgtagg gaaactctct cggtccagcc
     2461 cagtggtgac cacactgatg actcctgtgt tcctgttaat ggtgaacata tttttgtcag
     2521 ggagctcagg atcttggctg aggatggtgt aagcgatggc ggcattgtag gtgttcacat
     2581 catcgtccgc gtctgtggct gtgacctcca tcacagaggt tcctggaaga gcaccttcca
     2641 tgacagaccc cttaaagacc tcctgggtga attcgggctt gttgtcattc tgatcggtta
     2701 ccgtgatcaa aatctccatt ggatcctcaa ctgcattccc gttggatgac acagcgtgag
     2761 agaagagagt gtatgtggca atgcgttctc tatccagagg ctctgtcacc ttcagccatc
     2821 ctgtttctct ttcaataata aagacaccaa cagggggtgt gtcagctcct tggccagtga
     2881 tgctgtagaa aaccttgcct tctttgtctt tgttggattt gatctgaacc aggtttttag
     2941 gaaatgggcc tttttcattt tctgggcagc tgatgggagg aataacccag tctctcttct
     3001 gtcttctgag gccaggagag gagttgggaa atgtgagcaa ttctgcttgg attccagaaa
     3061 cggaggcctg atggggcggg gggcggtggt ggtgccccac tgtattcagc gtgactttgg
     3121 tggaaaactt tctgtaggtg gagtcccagg cgtagaccaa gaaatggatc tgtgggttat
     3181 gaaaccgtag aggccttttg actgtaatca caccatctgt gcccactttg aatcgggtgt
     3241 cgagggaaaa ataggctgtc ctttgtcgac cggtgcaatc ttcaaaattc actctgccca
     3301 ggacgcggcc tctctccagg tggcgccggg gcaccgtgaa cgtgtagctc tcggcgtcaa
     3361 agccagggtg gcagggctcc ggctcctggc agagccaaga ggagacctgc agcagcagca
     3421 gcagcgccga gaggctgcgg ctccaagggc ccatggcgat cgctagctgc aaaaagaaca
     3481 agtagcttgt attctatagt gtcacctaaa tgggcgatct gacggttcac taaaccagct
     3541 ctgcttatat aggcctccct ggcgcgccac tagttattaa tagtaatcaa ttacggggtc
     3601 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     3661 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     3721 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     3781 cttggcagta catcaagtgt atcatatgcc aagtccgccc cctattgacg tcaatgacgg
     3841 taaatggccc gcctggcatt atgcccagta catgacctta cgggactttc ctacttggca
     3901 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacaccaa
     3961 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     4021 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta ataaccccgc
     4081 cccgttgacg caaatgggcg gtaggcgtgt acggtgggag gtcaaatcga attctaatac
     4141 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa
     4201 ctaaacccag gaggcagatc atggagaaaa aaatcactgg atataccacc gttgatatat
     4261 cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata
     4321 accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca
     4381 agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaattcc
     4441 gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg
     4501 ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc
     4561 ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt
     4621 tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca
     4681 ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcactatgg
     4741 gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg
     4801 ccgtctgtga tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg
     4861 agtggcaggg cggggcgtaa gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     4921 caaccggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4981 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5041 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5101 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5161 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5221 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5281 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5341 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5401 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5461 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     5521 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5581 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatttcaa
     5641 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5701 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt atagtccgga
     5761 atcgtcacac aaaaaaccaa cacacagatg taatgaaaat aaagatattt tattgcggcc
     5821 atcgtgatgg ctagagttca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc
     5881 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc
     5941 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc
     6001 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag
     6061 gcatcgccgt gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg
     6121 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga
     6181 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
     6241 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
     6301 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
     6361 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg
     6421 gccagccacg atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg
     6481 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag
     6541 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga
     6601 gaacctgcgt gcaatccatc ttgttcaatc atggtggcat cgattcccgg tcaacatcca
     6661 gtacacctcc ggcaccaccg gcttccccaa gggcgccacg ctctcccacc acaacatcct
     6721 caacaacggc tactgggtgg gccgcacggt cggctacacg gagcaggacc gggtgtgtct
     6781 gccggtgccc ttctaccact gcttcggcat ggtcatgggc aacttgggcg ccacctccca
     6841 cgccgcctgc atcgtcatcc cggccccgtc cttcgagccg gcggcgacgc tggaggcggt
     6901 gcagcgggag aggtgcacgt ccctgtacgg cgtccccacg aagccgaatt ccaccacact
     6961 ggactagatg catgctcgac agtgtgatgg atatctgcag aattcggctt tcacgacacc
     7021 tgaaatggaa gaaaaaaact ttgaaccact gtctgaggct tgagaatgaa ccaagatcca
     7081 aactcaaaaa gggcaaattc caaggagaat tacatcaagt gccaagctgg cctaacttca
     7141 gtctccaccc actcagtgtg gggaaactcc atcgcataaa acccctcccc ccaacctaaa
     7201 gacgacgtac tccaaaagct cgagaactaa tcgaggtgcc tggacggcgc ccggtactcc
     7261 gtggagtcac atgaagcgac ggctgaggac ggaaaggccc ttttcctttg tgtgggtgac
     7321 tcacccgccc gctctcccga gcgccgcgtc ctccattttg agctgcctgc agcagggccg
     7381 ggaagcggcc atctttccgc tcacgcaact ggtgccgacc gggccagcct tgccgcccag
     7441 ggcggggcga tacacggcgg cgcgaggcca ggcaccagag caggccggcc agcttgagac
     7501 tacccccgtc cgattctcgg tggccgcgct cgcaggcccc gcctcgccga acatgtgcgc
     7561 tgggacgcac gggccccgtc gccgcccgcg gccccaaaaa ccgaaatacc agtgtgcaga
     7621 tcttggcccg catttacaag actatcttgc cagaaaaaaa gcgtcgcagc aggtcatcaa
     7681 aaattttaaa tggctagaga cttatcgaaa gcagcgagac aggcgcgaag gtgccaccag
     7741 attcgcacgc ggcggcccca gcgcccaggc caggcctcaa ctcaagcacg aggcgaaggg
     7801 gctccttaag cgcaaggcct cgaactctcc cacccacttc caacccgaag ctcgggatca
     7861 agaatcacgt actgcagcca ggtggaagta attcaaggca cgcaagggcc ataacccgta
     7921 aagaggccag gcccgcggga accacacacg gcacttacct gtgttctggc ggcaaacccg
     7981 ttgcgaaaaa gaacgttcac ggcgactact gcacttatat acggttctcc cccaccctcg
     8041 ggaaaaaggc ggagccagta cacgacatca ctttcccagt ttaccccgcg ccaccttctc
     8101 taggcaccgg ttcaattgcc gacccctccc cccaacttct cggggactgt gggcgatgtg
     8161 cgctctgccc actgacgggc accggagcc
//
