LOCUS       sequence-478476-        5769 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express XIAP-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 5769)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5769)
  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            "Unpublished"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: XIAP-HiBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5769
                     /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(2712..2730)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2767..2888
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2778..2797)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2832..2851
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3133..3490
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3133..3324
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3156..3176)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3341..3476
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3505..3552
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3584..4378
                     /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(3638..3657)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4248..4267
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4442..4490
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4729..5317)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4809..4828)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5433..5716
                     /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 ggctagcgat cgccatgact tttaacagtt
     1081 ttgaaggatc taaaacttgt gtacctgcag acatcaataa ggaagaagaa tttgtagaag
     1141 agtttaatag attaaaaact tttgctaatt ttccaagtgg tagtcctgtt tcagcatcaa
     1201 cactggcacg agcagggttt ctttatactg gtgaaggaga taccgtgcgg tgctttagtt
     1261 gtcatgcagc tgtagataga tggcaatatg gagactcagc agttggaaga cacaggaaag
     1321 tatccccaaa ttgcagattt atcaacggct tttatcttga aaatagtgcc acgcagtcta
     1381 caaattctgg tatccagaat ggtcagtaca aagttgaaaa ctatctggga agcagagatc
     1441 attttgcctt agacaggcca tctgagacac atgcagacta tcttttgaga actgggcagg
     1501 ttgtagatat atcagacacc atatacccga ggaaccctgc catgtatagt gaagaagcta
     1561 gattaaagtc ctttcagaac tggccagact atgctcacct aaccccaaga gagttagcaa
     1621 gtgctggact ctactacaca ggtattggtg accaagtgca gtgcttttgt tgtggtggaa
     1681 aactgaaaaa ttgggaacct tgtgatcgtg cctggtcaga acacaggcga cactttccta
     1741 attgcttctt tgttttgggc cggaatctta atattcgaag tgaatctgat gctgtgagtt
     1801 ctgataggaa tttcccaaat tcaacaaatc ttccaagaaa tccatccatg gcagattatg
     1861 aagcacggat ctttactttt gggacatgga tatactcagt taacaaggag cagcttgcaa
     1921 gagctggatt ttatgcttta ggtgaaggtg ataaagtaaa gtgctttcac tgtggaggag
     1981 ggctaactga ttggaagccc agtgaagacc cttgggaaca acatgctaaa tggtatccag
     2041 ggtgcaaata tctgttagaa cagaagggac aagaatatat aaacaatatt catttaactc
     2101 attcacttga ggagtgtctg gtaagaacta ctgagaaaac accatcacta actagaagaa
     2161 ttgatgatac catcttccaa aatcctatgg tacaagaagc tatacgaatg gggttcagtt
     2221 tcaaggacat taagaaaata atggaggaaa aaattcagat atctgggagc aactataaat
     2281 cacttgaggt tctggttgca gatctagtga atgctcagaa agacagtatg caagatgagt
     2341 caagtcagac ttcattacag aaagagatta gtactgaaga gcagctaagg cgcctgcaag
     2401 aggagaagct ttgcaaaatc tgtatggata gaaatattgc tatcgttttt gttccttgtg
     2461 gacatctagt cacttgtaaa caatgtgctg aagcagttga caagtgtccc atgtgctaca
     2521 cagtcattac tttcaagcaa aaaattttta tgtctgtttc tcaaggcagt tcaggtggct
     2581 cgagcggtgt gagcggctgg cggctgttca agaagattag ctaaggccgc gactctagag
     2641 tcgacctgca ggcatgcaag ctgatccggc tgctaacaaa gcccgaaagg aagctgagtt
     2701 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcggccg cttcgagcag
     2761 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     2821 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     2881 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg
     2941 aggttttttt aagcaagtaa aacctctaca aatgtggtaa aatcgaattt taacaaaata
     3001 ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3061 accgcatacg cggatctgcg cagcaccatg gcctgaaata acctctgaaa gaggaacttg
     3121 gttaggtacc ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     3181 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3241 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3301 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3361 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3421 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3481 cttttgcaaa aagcttaatt aactgttgac aattaatcat cggcatagta tatcggcata
     3541 gtataatacg acaaggtgag gaactaaacc caggaggcag atcatgattg aacaagatgg
     3601 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     3661 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     3721 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     3781 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     3841 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     3901 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     3961 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4021 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4081 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     4141 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4201 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4261 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4321 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     4381 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgatggccg
     4441 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagc
     4501 gataaggatc ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt
     4561 catccggggt cagcaccgtt tctgcggact ggctttctac ccggtatcag ctcactcaaa
     4621 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4681 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4741 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4801 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4861 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4921 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4981 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5041 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5101 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5161 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5221 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5281 caagcagcag attacgcgca gaaaaaaagg atttcaagaa gatcctttga tcttttctac
     5341 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5401 aaaaaggatc ttcacctaga tccttttata gtccggaaat acaggaacgc acgctggatg
     5461 gcccttcgct gggatggtga aaccatgaaa aatggcagct tcagtggatt aagtgggggt
     5521 aatgtggcct gtaccctctg gttgcatagg tattcatacg gttaaaattt atcaggcgcg
     5581 attgcggcag tttttcgggt ggtttgttgc catttttacc tgtctgctgc cgtgatcgcg
     5641 ctgaacgcgt tttagcggtg cgtacaatta agggattatg gtaaatccac ttactgtctg
     5701 ccctcgtagc catcgagata aaccgcagta ctccggccac gatgcgtccg gcgtagagga
     5761 tcgagatct
//
