LOCUS       sequence-475107-        8000 bp    DNA     circular SYN 24-JUN-2025
DEFINITION  Express EPHA2-HaloTag-HiBiT 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 8000)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 8000)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 24, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: EPHA2-HaloTag-HiBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..8000
                     /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"
     CDS             4014..4034
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             4050..4934
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     primer_bind     complement(5079..5097)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    5134..5255
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5145..5164)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5199..5218
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5500..5857
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5500..5691
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5523..5543)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5708..5843
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5872..5919
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5951..6745
                     /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(6005..6024)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6615..6634
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6809..6857
                     /note="synthetic polyadenylation signal"
     rep_origin      7014..7559
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     7460..7479
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7676..7693
                     /label=L4440
                     /note="L4440 vector, forward primer"
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 ggctagcgat cgccatggag ctccaggcag
     1081 cccgcgcctg cttcgccctg ctgtggggct gtgcgctggc cgcggccgcg gcggcgcagg
     1141 gcaaggaagt ggtactgctg gactttgctg cagctggagg ggagctcggc tggctcacac
     1201 acccgtatgg caaagggtgg gacctgatgc agaacatcat gaatgacatg ccgatctaca
     1261 tgtactccgt gtgcaacgtg atgtctggcg accaggacaa ctggctccgc accaactggg
     1321 tgtaccgagg agaggctgag cgtatcttca ttgagctcaa gtttactgta cgtgactgca
     1381 acagcttccc tggtggcgcc agctcctgca aggagacttt caacctctac tatgccgagt
     1441 cggacctgga ctacggcacc aacttccaga agcgcctgtt caccaagatt gacaccattg
     1501 cgcccgatga gatcaccgtc agcagcgact tcgaggcacg ccacgtgaag ctgaacgtgg
     1561 aggagcgctc cgtggggccg ctcacccgca aaggcttcta cctggccttc caggatatcg
     1621 gtgcctgtgt ggcgctgctc tccgtccgtg tctactacaa gaagtgcccc gagctgctgc
     1681 agggcctggc ccacttccct gagaccatcg ccggctctga tgcaccttcc ctggccactg
     1741 tggccggcac ctgtgtggac catgccgtgg tgccaccggg gggtgaagag ccccgtatgc
     1801 actgtgcagt ggatggcgag tggctggtgc ccattgggca gtgcctgtgc caggcaggct
     1861 acgagaaggt ggaggatgcc tgccaggcct gctcgcctgg attttttaag tttgaggcat
     1921 ctgagagccc ctgcttggag tgccctgagc acacgctgcc atcccctgag ggtgccacct
     1981 cctgcgagtg tgaggaaggc ttcttccggg cacctcagga cccagcgtcg atgccttgca
     2041 cacgaccccc ctccgcccca cactacctca cagccgtggg catgggtgcc aaggtggagc
     2101 tgcgctggac gccccctcag gacagcgggg gccgcgagga cattgtctac agcgtcacct
     2161 gcgaacagtg ctggcccgag tctggggaat gcgggccgtg tgaggccagt gtgcgctact
     2221 cggagcctcc tcacggactg acccgcacca gtgtgacagt gagcgacctg gagccccaca
     2281 tgaactacac cttcaccgtg gaggcccgca atggcgtctc aggcctggta accagccgca
     2341 gcttccgtac tgccagtgtc agcatcaacc agacagagcc ccccaaggtg aggctggagg
     2401 gccgcagcac cacctcgctt agcgtctcct ggagcatccc cccgccgcag cagagccgag
     2461 tgtggaagta cgaggtcact taccgcaaga agggagactc caacagctac aatgtgcgcc
     2521 gcaccgaggg tttctccgtg accctggacg acctggcccc agacaccacc tacctggtcc
     2581 aggtgcaggc actgacgcag gagggccagg gggccggcag caaggtgcac gaattccaga
     2641 cgctgtcccc ggagggatct ggcaacttgg cggtgattgg cggcgtggct gtcggtgtgg
     2701 tcctgcttct ggtgctggca ggagttggct tctttatcca ccgcaggagg aagaaccagc
     2761 gtgcccgcca gtccccggag gacgtttact tctccaagtc agaacaactg aagcccctga
     2821 agacatacgt ggacccccac acatatgagg accccaacca ggctgtgttg aagttcacta
     2881 ccgagatcca tccatcctgt gtcactcggc agaaggtgat cggagcagga gagtttgggg
     2941 aggtgtacaa gggcatgctg aagacatcct cggggaagaa ggaggtgccg gtggccatca
     3001 agacgctgaa agccggctac acagagaagc agcgagtgga cttcctcggc gaggccggca
     3061 tcatgggcca gttcagccac cacaacatca tccgcctaga gggcgtcatc tccaaataca
     3121 agcccatgat gatcatcact gagtacatgg agaatggggc cctggacaag ttccttcggg
     3181 agaaggatgg cgagttcagc gtgctgcagc tggtgggcat gctgcggggc atcgcagctg
     3241 gcatgaagta cctggccaac atgaactatg tgcaccgtga cctggctgcc cgcaacatcc
     3301 tcgtcaacag caacctggtc tgcaaggtgt ctgactttgg cctgtcccgc gtgctggagg
     3361 acgaccccga ggccacctac accaccagtg gcggcaagat ccccatccgc tggaccgccc
     3421 cggaggccat ttcctaccgg aagttcacct ctgccagcga cgtgtggagc tttggcattg
     3481 tcatgtggga ggtgatgacc tatggcgagc ggccctactg ggagttgtcc aaccacgagg
     3541 tgatgaaagc catcaatgat ggcttccggc tccccacacc catggactgc ccctccgcca
     3601 tctaccagct catgatgcag tgctggcagc aggagcgtgc ccgccgcccc aagttcgctg
     3661 acatcgtcag catcctggac aagctcattc gtgcccctga ctccctcaag accctggctg
     3721 actttgaccc ccgcgtgtct atccggctcc ccagcacgag cggctcggag ggggtgccct
     3781 tccgcacggt gtccgagtgg ctggagtcca tcaagatgca gcagtatacg gagcacttca
     3841 tggcggccgg ctacactgcc atcgagaagg tggtgcagat gaccaacgac gacatcaaga
     3901 ggattggggt gcggctgccc ggccaccaga agcgcatcgc ctacagcctg ctgggactca
     3961 aggaccaggt gaacactgtg gggatcccca tcgtttctct cgagccaacc actgaggatc
     4021 tgtactttca gagcgataac gatggatccg aaatcggtac tggctttcca ttcgaccccc
     4081 attatgtgga agtcctgggc gagcgcatgc actacgtcga tgttggtccg cgcgatggca
     4141 cccctgtgct gttcctgcac ggtaacccga cctcctccta cgtgtggcgc aacatcatcc
     4201 cgcatgttgc accgacccat cgctgcattg ctccagacct gatcggtatg ggcaaatccg
     4261 acaaaccaga cctgggttat ttcttcgacg accacgtccg cttcatggat gccttcatcg
     4321 aagccctggg tctggaagag gtcgtcctgg tcattcacga ctggggctcc gctctgggtt
     4381 tccactgggc caagcgcaat ccagagcgcg tcaaaggtat tgcatttatg gagttcatcc
     4441 gccctatccc gacctgggac gaatggccag aatttgcccg cgagaccttc caggccttcc
     4501 gcaccaccga cgtcggccgc aagctgatca tcgatcagaa cgtttttatc gagggtacgc
     4561 tgccgatggg tgtcgtccgc ccgctgactg aagtcgagat ggaccattac cgcgagccgt
     4621 tcctgaatcc tgttgaccgc gagccactgt ggcgcttccc aaacgagctg ccaatcgccg
     4681 gtgagccagc gaacatcgtc gcgctggtcg aagaatacat ggactggctg caccagtccc
     4741 ctgtcccgaa gctgctgttc tggggcaccc caggcgttct gatcccaccg gccgaagccg
     4801 ctcgcctggc caaaagcctg cctaactgca aggctgtgga catcggcccg ggtctgaatc
     4861 tgctgcaaga agacaacccg gacctgatcg gcagcgagat cgcgcgctgg ctgtctactc
     4921 tggagatttc cggtggcagt tcaggtggct cgagcggtgt gagcggctgg cggctgttca
     4981 agaagattag ctaatagaat tctagagtcg acctgcaggc atgcaagctg atccggctgc
     5041 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     5101 accccttggg gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa
     5161 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     5221 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     5281 tgtttcaggt tcagggggag atgtgggagg tttttttaag caagtaaaac ctctacaaat
     5341 gtggtaaaat cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc
     5401 tccttacgca tctgtgcggt atttcacacc gcatacgcgg atctgcgcag caccatggcc
     5461 tgaaataacc tctgaaagag gaacttggtt aggtaccttc tgaggcggaa agaaccagct
     5521 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5581 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     5641 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     5701 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     5761 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     5821 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttaattaac tgttgacaat
     5881 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaacccag
     5941 gaggcagatc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     6001 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     6061 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     6121 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     6181 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     6241 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     6301 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     6361 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     6421 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     6481 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     6541 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     6601 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     6661 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     6721 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     6781 acgcccaacc tgccatcacg atggccgcaa taaaatatct ttattttcat tacatctgtg
     6841 tgttggtttt ttgtgtgaat cgatagcgat aaggatccgc tcatgcgatg ataagctgtc
     6901 aaacatgaga attacaactt atatcgtatg gggctgactt caggtgctac atttgaagag
     6961 ataaattgca ctgaaatcta gaaatatttt atctgattaa taagatgatc ttcttgagat
     7021 cgttttggtc tgcgcgtaat ctcttgctct gaaaacgaaa aaaccgcctt gcagggcggt
     7081 ttttcgaagg ttctctgagc taccaactct ttgaaccgag gtaactggct tggaggagcg
     7141 cagtcaccaa aacttgtcct ttcagtttag ccttaaccgg cgcatgactt caagactaac
     7201 tcctctaaat caattaccag tggctgctgc cagtggtgct tttgcatgtc tttccgggtt
     7261 ggactcaaga cgatagttac cggataaggc gcagcggtcg gactgaacgg ggggttcgtg
     7321 catacagtcc agcttggagc gaactgccta cccggaactg agtgtcaggc gtggaatgag
     7381 acaaacgcgg ccataacagc ggaatgacac cggtaaaccg aaaggcagga acaggagagc
     7441 gcacgaggga gccgccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     7501 accactgatt tgagcgtcag atttcgtgat gcttgtcagg ggggcggagc ctatggaaaa
     7561 acggctttgc cgcggccctc tcacttccct gttaagtatc ttcctggcat cttccaggaa
     7621 atctccgccc cgttcgtaag ccatttccgc tcgccgcagt cgaacgaccg agcgtagcga
     7681 gtcagtgagc gaggaagcgg aatatatcct gtatcacata ttctgctgac gcaccggtgc
     7741 agcctttttt ctcctgccac atgaagcact tcactgacac cctcatcagt gccaacatag
     7801 taagccagta tacactccgc tagcgctgat gtccggcggt gcttttgccg ttacgcacca
     7861 ccccgtcatg agattacgcg ttttagcggt gcgtacaatt aagggattat ggtaaatcca
     7921 cttactgtct gccctcgtag ccatcgagat aaaccgcagt actccggcca cgatgcgtcc
     7981 ggcgtagagg atcgagatct
//
