LOCUS       sequence-478491-        6576 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express ZER1-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 6576)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6576)
  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            "Unpublished"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: ZER1-HiBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6576
                     /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(3519..3537)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3574..3695
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3585..3604)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3639..3658
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3940..4297
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3940..4131
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3963..3983)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4148..4283
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4312..4359
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4391..5185
                     /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(4445..4464)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5055..5074
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5249..5297
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5536..6124)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5616..5635)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6240..6523
                     /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 cgccatggcg tccgacactc
     1081 ccgagtcgct gatggccctc tgtactgact tctgcttgcg caacctggat ggcaccctgg
     1141 gctacctgct ggacaaggag accctgcggc tacatccgga catcttcttg cccagcgaga
     1201 tctgtgaccg gctcgtcaat gagtatgtgg agctggtgaa cgctgcctgt aacttcgagc
     1261 cacacgagag cttcttcagc ctcttttcgg acccccgcag cacccgcctc acgcggatcc
     1321 acctccgtga ggacctggtg caggaccagg acctggaggc catccgcaag caggacctgg
     1381 tggagctgta cctgactaac tgcgagaagc tgtccgccaa gagcctgcag acactgagga
     1441 gcttcagcca caccctggtg tccttgagcc tcttcggctg tacaaacatt ttctatgagg
     1501 aggagaaccc agggggctgt gaagatgagt acctcgtcaa ccccacctgc caggtgctgg
     1561 ttaaggattt caccttcgag ggcttcagcc gcctccgctt cctcaacttg ggccgcatga
     1621 ttgattgggt ccctgtggag tccctgctgc ggccgcttaa ctccctggct gccttggacc
     1681 tctcaggcat tcagacgagc gacgcagcct tcctcaccca gtggaaagac agcctggtgt
     1741 ccctcgtcct ctacaacatg gacctgtccg acgaccacat ccgggtcatc gtgcagctgc
     1801 acaagctgcg acacctggac atctcccgag accgcctctc cagctactac aagttcaagc
     1861 tgactcggga ggtgctgagc ctctttgtgc agaagctggg gaacctaatg tccctggaca
     1921 tctctggcca catgatccta gagaactgca gcatctccaa gatggaagag gaagcggggc
     1981 agaccagcat tgagccttcc aagagcagca tcataccttt ccgggctctg aagaggccgc
     2041 tgcagttcct cgggctcttt gagaactctc tgtgccgcct cacgcacatt ccagcctaca
     2101 aagtaagtgg tgacaaaaac gaagagcagg tgctgaatgc catcgaggcc tacacggagc
     2161 accggcctga gatcacctcg cgggccatca acttgctttt tgacatcgcc cgcatcgagc
     2221 gttgcaacca gctgctgcgg gccctgaagc tggtcatcac ggccctcaag tgccacaaat
     2281 atgacaggaa cattcaagtg acaggcagcg ccgctctctt ctacctaaca aattccgagt
     2341 accgctcaga gcagagtgtg aagctgcgcc ggcaggttat ccaggtggtg ctgaatggca
     2401 tggaatccta ccaggaggtg acggtgcagc ggaactgctg cctgacgctc tgcaacttca
     2461 gcatccccga ggagctggaa ttccagtacc gccgggtcaa cgagctcctg ctcagcatcc
     2521 tcaaccccac gcggcaggac gagtctatcc agcggatcgc cgtgcacctg tgcaatgccc
     2581 tggtctgcca ggtagacaac gaccacaagg aggccgtggg caagatgggc tttgtcgtga
     2641 ccatgctgaa gctgattcag aagaagctgc tggacaagac atgtgaccag gtcatggagt
     2701 tctcctggag tgccctgtgg aacatcacag atgaaactcc tgacaactgc gagatgttcc
     2761 tcaatttcaa cggcatgaag ctcttcctgg actgcctgaa ggaattccca gagaagcagg
     2821 aactgcatag gaatatgcta ggacttttgg ggaatgtggc agaagtgaag gagctgaggc
     2881 ctcaactaat gacttcccag ttcatcagcg tcttcagcaa cctgttggag agcaaggccg
     2941 atgggatcga ggtttcctac aatgcctgcg gcgtcctctc ccacatcatg tttgatggac
     3001 ccgaggcctg gggcgtctgt gagccccagc gtgaggaggt ggaggaacgc atgtgggctg
     3061 ccatccagag ctgggacata aactctcgga gaaacatcaa ttacaggtca tttgaaccaa
     3121 ttctccgcct ccttccccag ggaatctctc ctgtcagcca gcactgggca acctgggccc
     3181 tgtataacct cgtgtctgtc tacccggaca agtactgccc tctgctgatc aaagaagggg
     3241 ggatgcccct tctgagggac ataattaaga tggcgaccgc acggcaggag accaaggaaa
     3301 tggcccgcaa ggtgattgag cactgcagta actttaaaga ggagaacatg gacacgtcta
     3361 gagtttctca aggcagttca ggtggctcga gcggtgtgag cggctggcgg ctgttcaaga
     3421 agattagcta aggccgcgac tctagagtcg acctgcaggc atgcaagctg atccggctgc
     3481 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     3541 accccttggg gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa
     3601 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     3661 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     3721 tgtttcaggt tcagggggag atgtgggagg tttttttaag caagtaaaac ctctacaaat
     3781 gtggtaaaat cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc
     3841 tccttacgca tctgtgcggt atttcacacc gcatacgcgg atctgcgcag caccatggcc
     3901 tgaaataacc tctgaaagag gaacttggtt aggtaccttc tgaggcggaa agaaccagct
     3961 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4021 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4081 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4141 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4201 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4261 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttaattaac tgttgacaat
     4321 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaacccag
     4381 gaggcagatc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4441 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4501 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4561 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4621 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4681 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4741 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4801 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4861 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     4921 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4981 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5041 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5101 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5161 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     5221 acgcccaacc tgccatcacg atggccgcaa taaaatatct ttattttcat tacatctgtg
     5281 tgttggtttt ttgtgtgaat cgatagcgat aaggatcctc tttgcgcttg cgttttccct
     5341 tgtccagata gcccagtagc tgacattcat ccggggtcag caccgtttct gcggactggc
     5401 tttctacccg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5461 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5521 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5581 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5641 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5701 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5761 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5821 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5881 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5941 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     6001 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6061 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt
     6121 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6181 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttatagtc
     6241 cggaaataca ggaacgcacg ctggatggcc cttcgctggg atggtgaaac catgaaaaat
     6301 ggcagcttca gtggattaag tgggggtaat gtggcctgta ccctctggtt gcataggtat
     6361 tcatacggtt aaaatttatc aggcgcgatt gcggcagttt ttcgggtggt ttgttgccat
     6421 ttttacctgt ctgctgccgt gatcgcgctg aacgcgtttt agcggtgcgt acaattaagg
     6481 gattatggta aatccactta ctgtctgccc tcgtagccat cgagataaac cgcagtactc
     6541 cggccacgat gcgtccggcg tagaggatcg agatct
//
