LOCUS       sequence-478472-        5224 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express HiBiT-TREX1 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 5224)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5224)
  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            "Unpublished"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-TREX1 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        338..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1157..1176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1157..1175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2287..2408
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2298..2317)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2523..2556)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2566..2613
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2645..3439
                     /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(2699..2718)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3309..3328
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3597..4185)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3677..3696)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4311..4359
                     /note="synthetic polyadenylation signal"
     CDS             complement(4383..4781)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4778..4787
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4812..5169)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4826..4961
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4978..5169
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5126..5146
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaatctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact ataggggtta gcgatcgcca tggtgagcgg
     1201 ctggcggctg ttcaagaaga ttagcgggag ctccggtggc tcgagcggtg gatctcaggc
     1261 gttgccccct ggccctatgc aaactctcat attttttgac atggaagcga ccggactccc
     1321 ctttagccag ccaaaagtga ctgaactgtg ccttctcgcc gtgcatcgat gcgctctgga
     1381 atccccacca acgagtcagg ggccacctcc gacagtaccg ccgcccccta gagttgtcga
     1441 caagctttct ctgtgcgtcg cacctggaaa agcttgtagc ccggccgcct ctgagattac
     1501 aggactttct accgcagtat tggctgcgca cggtcgccag tgcttcgacg acaatttggc
     1561 aaatctcctg ttggcttttc ttagacgcca gccccaaccc tggtgtctgg ttgctcacaa
     1621 cggtgaccgg tacgattttc cgctgctcca agcggaattg gcgatgctgg gtcttacttc
     1681 cgccctcgac ggggctttct gtgtcgactc cataacggcc ctgaaagctc ttgagcgagc
     1741 gtcatcacca agcgagcacg gccccagaaa atcttattca ctgggatcca tatatactcg
     1801 cctgtatggg cagagtcccc cagatagcca caccgcagaa ggtgatgtct tggctctcct
     1861 cagtatttgc caatggagac cccaggcgtt gctcagatgg gtggatgccc atgcaagacc
     1921 ctttgggacc attcggccga tgtatggggt gactgcttcc gctcgcacta agcccaggcc
     1981 gagcgcagtt actaccacgg cgcatctggc aacaacccgg aacacttccc cttcactggg
     2041 cgagagccgg ggtacgaaag atctccctcc cgtaaaagac cccggtgcgc tttcaaggga
     2101 aggtcttttg gccccactcg gccttttggc tattttgacg ctggcagtag ccacgttgta
     2161 cggcttgtct ctggcgacac ccggagagta attctagagg tcaagcttcg ggggtttaaa
     2221 cctgcaggca tgcaagctga tttggctgct gccaccgctg agcaaggccg cttcgagcag
     2281 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     2341 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     2401 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg
     2461 aggttttttt aagcaagtaa aacctctaca aatgtggtaa aatcttaatt aagacttcta
     2521 atataacttc gtatagcata cattatacga agttatggtt ccactgttga caattaatca
     2581 tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac ccaggaggca
     2641 gatcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     2701 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     2761 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     2821 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     2881 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     2941 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3001 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     3061 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     3121 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     3181 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     3241 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     3301 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     3361 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     3421 tcttgacgag ttcttctgat tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct
     3481 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3541 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3601 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3661 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3721 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3781 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3841 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3901 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3961 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4021 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4081 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4141 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc
     4201 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4261 agattatcaa aaaggatctt cacctagatc cttttatagt ccggaatcgt cacacaaaaa
     4321 accaacacac agatgtaatg aaaataaaga tattttattg cggccatcgt gatggctaga
     4381 gtttagccct cccacacata accagagggc agcaattcac gaatcccaac tgccgtcggc
     4441 tgtccatcac tgtccttcac tatggctttg atcccaggat gcagatcgag aagcacctgt
     4501 cggcaccgtc cgcaggggct caagatgccc ctgttctcat ttccgatcgc gacgatacaa
     4561 gtcaggttgc cagctgccgc agcagcagca gtgcccagca ccacgagttc tgcacaaggt
     4621 cccccagtaa aatgatatac attgacacca gtgaagatgc ggccgtcgct agagagagct
     4681 gcgctggcga cgctgtagtc ttcagagatg gggatgctgt tgattgtagc cgttgctctt
     4741 tcaatgaggg tggattcttc ttgagacaaa ggcttggcca tggtggcgct agtgtcagaa
     4801 gaatcgagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat
     4861 agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg
     4921 ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg
     4981 actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
     5041 ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct
     5101 gctggggagc ctggggactt tccacaccct aactgacaca cattccacag ctggttcttt
     5161 ccgcctcaga aggtacctaa ccaagttcct ctttcagagg ttatttcagg ccatggtgct
     5221 gcgc
//
