LOCUS       sequence-478469-        7946 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express HiBiT-EGFR 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 7946)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7946)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: HiBiT-EGFR Fusion Vector
FEATURES             Location/Qualifiers
     source          1..7946
                     /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(4889..4907)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4944..5065
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4955..4974)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5009..5028
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5310..5667
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5310..5501
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5333..5353)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5518..5653
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5682..5729
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5761..6555
                     /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(5815..5834)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6425..6444
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6619..6667
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(6906..7494)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6986..7005)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7610..7893
                     /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 caccatgaac tccttctcca
     1081 caagcgcctt cggtccagtt gccttctccc tgggcctgct cctggtgttg cctgctgcct
     1141 tccctgcccc agtgagcggc tggcggctgt tcaagaagat tagcgggagc tccggtggct
     1201 cgagcggtgc gatcgccctg gaggaaaaga aagtttgcca aggcacgagt aacaagctca
     1261 cgcagttggg cacttttgaa gatcattttc tcagcctcca gaggatgttc aataactgtg
     1321 aggtggtcct tgggaatttg gaaattacct atgtgcagag gaattatgat ctttccttct
     1381 taaagaccat ccaggaggtg gctggttatg tcctcattgc cctcaacaca gtggagcgaa
     1441 ttcctttgga aaacctgcag atcatcagag gaaatatgta ctacgaaaat tcctatgcct
     1501 tagcagtctt atctaactat gatgcaaata aaaccggact gaaggagctg cccatgagaa
     1561 atttacagga aatcctgcat ggcgccgtgc ggttcagcaa caaccctgcc ctgtgcaacg
     1621 tggagagcat ccagtggcgg gacatagtca gcagtgactt tctcagcaac atgtcgatgg
     1681 acttccagaa ccacctgggc agctgccaaa agtgtgatcc aagctgtccc aatgggagct
     1741 gctggggtgc aggagaggag aactgccaga aactgaccaa aatcatctgt gcccagcagt
     1801 gctccgggcg ctgccgtggc aagtccccca gtgactgctg ccacaaccag tgtgctgcag
     1861 gctgcacagg cccccgggag agcgactgcc tggtctgccg caaattccga gacgaagcca
     1921 cgtgcaagga cacctgcccc ccactcatgc tctacaaccc caccacgtac cagatggatg
     1981 tgaaccccga gggcaaatac agctttggtg ccacctgcgt gaagaagtgt ccccgtaatt
     2041 atgtggtgac agatcacggc tcgtgcgtcc gagcctgtgg ggccgacagc tatgagatgg
     2101 aggaagacgg cgtccgcaag tgtaagaagt gcgaagggcc ttgccgcaaa gtgtgtaacg
     2161 gaataggtat tggtgaattt aaagactcac tctccataaa tgctacgaat attaaacact
     2221 tcaaaaactg cacctccatc agtggcgatc tccacatcct gccggtggca tttaggggtg
     2281 actccttcac acatactcct cctctggatc cacaggaact ggatattctg aaaaccgtaa
     2341 aggaaatcac agggtttttg ctgattcagg cttggcctga aaacaggacg gacctccatg
     2401 cctttgagaa cctagaaatc atacgcggca ggaccaagca acatggtcag ttttctcttg
     2461 cagtcgtcag cctgaacata acatccttgg gattacgctc cctcaaggag ataagtgatg
     2521 gagatgtgat aatttcagga aacaaaaatt tgtgctatgc aaatacaata aactggaaaa
     2581 aactgtttgg gacctccggt cagaaaacca aaattataag caacagaggt gaaaacagct
     2641 gcaaggccac aggccaggtc tgccatgcct tgtgctcccc cgagggctgc tggggcccgg
     2701 agcccaggga ctgcgtctct tgccggaatg tcagccgagg cagggaatgc gtggacaagt
     2761 gcaaccttct ggagggtgag ccaagggagt ttgtggagaa ctctgagtgc atacagtgcc
     2821 acccagagtg cctgcctcag gccatgaaca tcacctgcac aggacgggga ccagacaact
     2881 gtatccagtg tgcccactac attgacggcc cccactgcgt caagacctgc ccggcaggag
     2941 tcatgggaga aaacaacacc ctggtctgga agtacgcaga cgccggccat gtgtgccacc
     3001 tgtgccatcc aaactgcacc tacggatgca cagggccagg tcttgaaggc tgtccaacga
     3061 atgggcctaa gatcccgtcc atcgccactg ggatggtggg ggccctcctc ttgctgctgg
     3121 tggtggccct ggggatcggc ctcttcatgc gaaggcgcca catcgttcgg aagcgcacgc
     3181 tgcggaggct gctgcaggag agggagcttg tggagcctct tacacccagt ggagaagctc
     3241 ccaaccaagc tctcttgagg atcttgaagg aaactgaatt caaaaagatc aaagtgctgg
     3301 gctccggtgc gttcggcacg gtgtataagg gactctggat cccagaaggt gagaaagtta
     3361 aaattcccgt cgctatcaag gaattaagag aagcaacatc tccgaaagcc aacaaggaaa
     3421 tcctcgatga agcctacgtg atggccagcg tggacaaccc ccacgtgtgc cgcctgctgg
     3481 gcatctgcct cacctccacc gtgcagctca tcacgcagct catgcccttc ggctgcctcc
     3541 tggactatgt ccgggaacac aaagacaata ttggctccca gtacctgctc aactggtgtg
     3601 tgcagatcgc aaagggcatg aactacttgg aggaccgtcg cttggtgcac cgcgacctgg
     3661 cagccaggaa cgtactggtg aaaacaccgc agcatgtcaa gatcacagat tttgggctgg
     3721 ccaaactgct gggtgcggaa gagaaagaat accatgcaga aggaggcaaa gtgcctatca
     3781 agtggatggc attggaatca attttacaca gaatctatac ccaccagagt gatgtctgga
     3841 gctacggggt gaccgtttgg gagttgatga cctttggatc caagccatat gacggaatcc
     3901 ctgccagcga gatctcctcc atcctggaga aaggagaacg cctccctcag ccacccatat
     3961 gtaccatcga tgtctacatg atcatggtca agtgctggat gatagacgca gatagtcgcc
     4021 caaagttccg tgagttgatc atcgaattct ccaaaatggc ccgagacccc cagcgctacc
     4081 ttgtcattca gggggatgaa agaatgcatt tgccaagtcc tacagactcc aacttctacc
     4141 gtgccctgat ggatgaagaa gacatggacg acgtggtgga tgccgacgag tacctcatcc
     4201 cacagcaggg cttcttcagc agcccctcca cgtcacggac tcccctcctg agctctctga
     4261 gtgcaaccag caacaattcc accgtggctt gcattgatag aaatgggctg caaagctgtc
     4321 ccatcaagga agacagcttc ttgcagcgat acagctcaga ccccacaggc gccttgactg
     4381 aggacagcat agacgacacc ttcctcccag tgcctgaata cataaaccag tccgttccca
     4441 aaaggcccgc tggctctgtg cagaatcctg tctatcacaa tcagcctctg aaccccgcgc
     4501 ccagcagaga cccacactac caggaccccc acagcactgc agtgggcaac cccgagtatc
     4561 tcaacactgt ccagcccacc tgtgtcaaca gcacattcga cagccctgcc cactgggccc
     4621 agaaaggcag ccaccaaatt agcctggaca accctgacta ccagcaggac ttctttccca
     4681 aggaagccaa gccaaatggc atctttaagg gctccacagc tgaaaatgca gaatacctaa
     4741 gggtcgcgcc acaaagcagt gaatttattg gagcagttta aacgaattcg ggctcggtac
     4801 ccggggatcc tctagagtcg acctgcaggc atgcaagctg atccggctgc taacaaagcc
     4861 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     4921 gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     4981 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     5041 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     5101 tcagggggag atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat
     5161 cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca
     5221 tctgtgcggt atttcacacc gcatacgcgg atctgcgcag caccatggcc tgaaataacc
     5281 tctgaaagag gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg
     5341 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5401 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     5461 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     5521 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     5581 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     5641 ttttttggag gcctaggctt ttgcaaaaag cttaattaac tgttgacaat taatcatcgg
     5701 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaacccag gaggcagatc
     5761 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5821 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5881 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5941 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     6001 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     6061 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     6121 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     6181 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     6241 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     6301 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     6361 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     6421 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     6481 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     6541 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     6601 tgccatcacg atggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt
     6661 ttgtgtgaat cgatagcgat aaggatcctc tttgcgcttg cgttttccct tgtccagata
     6721 gcccagtagc tgacattcat ccggggtcag caccgtttct gcggactggc tttctacccg
     6781 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     6841 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6901 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6961 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     7021 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     7081 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     7141 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     7201 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     7261 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     7321 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     7381 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     7441 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat
     7501 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     7561 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttatagtc cggaaataca
     7621 ggaacgcacg ctggatggcc cttcgctggg atggtgaaac catgaaaaat ggcagcttca
     7681 gtggattaag tgggggtaat gtggcctgta ccctctggtt gcataggtat tcatacggtt
     7741 aaaatttatc aggcgcgatt gcggcagttt ttcgggtggt ttgttgccat ttttacctgt
     7801 ctgctgccgt gatcgcgctg aacgcgtttt agcggtgcgt acaattaagg gattatggta
     7861 aatccactta ctgtctgccc tcgtagccat cgagataaac cgcagtactc cggccacgat
     7921 gcgtccggcg tagaggatcg agatct
//
