LOCUS       sequence-478480-        6120 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express NR1D1-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 6120)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6120)
  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            "Unpublished"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: NR1D1-HiBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6120
                     /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(3063..3081)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3118..3239
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3129..3148)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3183..3202
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3484..3841
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3484..3675
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3507..3527)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3692..3827
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3856..3903
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3935..4729
                     /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(3989..4008)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4599..4618
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4793..4841
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5080..5668)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5160..5179)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5784..6067
                     /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 cgccatgacg accctggact
     1081 ccaacaacaa cacaggtggc gtcatcacct acattggctc cagtggctcc tccccaagcc
     1141 gcaccagccc tgaatccctc tatagtgaca actccaatgg cagcttccag tccctgaccc
     1201 aaggctgtcc cacctacttc ccaccatccc ccactggctc cctcacccaa gacccggctc
     1261 gctcctttgg gagcattcca cccagcctga gtgatgacgg ctccccttct tcctcatctt
     1321 cctcgtcgtc atcctcctcc tccttctata atgggagccc ccctgggagt ctacaagtgg
     1381 ccatggagga cagcagccga gtgtccccca gcaagagcac cagcaacatc accaagctga
     1441 atggcatggt gttactgtgt aaagtgtgtg gggacgttgc ctcgggcttc cactacggtg
     1501 tgcacgcctg cgagggctgc aagggctttt tccgtcggag catccagcag aacatccagt
     1561 acaaaaggtg tctgaagaat gagaattgct ccatcgtccg catcaatcgc aaccgctgcc
     1621 agcaatgtcg cttcaagaag tgtctctctg tgggcatgtc tcgagacgct gtgcgttttg
     1681 ggcgcatccc caaacgagag aagcagcgga tgcttgctga gatgcagagt gccatgaacc
     1741 tggccaacaa ccagttgagc agccagtgcc cgctggagac ttcacccacc cagcacccca
     1801 ccccaggccc catgggcccc tcgccacccc ctgctccggt cccctcaccc ctggtgggct
     1861 tctcccagtt tccacaacag ctgacgcctc ccagatcccc aagccctgag cccacagtgg
     1921 aggatgtgat atcccaggtg gcccgggccc atcgagagat cttcacctac gcccatgaca
     1981 agctgggcag ctcacctggc aacttcaatg ccaaccatgc atcaggtagc cctccagcca
     2041 ccaccccaca tcgctgggaa aatcagggct gcccacctgc ccccaatgac aacaacacct
     2101 tggctgccca gcgtcataac gaggccctaa atggtctgcg ccaggctccc tcctcctacc
     2161 ctcccacctg gcctcctggc cctgcacacc acagctgcca ccagtccaac agcaacgggc
     2221 accgtctatg ccccacccac gtgtatgcag ccccagaagg caaggcacct gccaacagtc
     2281 cccggcaggg caactcaaag aatgttctgc tggcatgtcc tatgaacatg tacccgcatg
     2341 gacgcagtgg gcgaacggtg caggagatct gggaggattt ctccatgagc ttcacgcccg
     2401 ctgtgcggga ggtggtagag tttgccaaac acatcccggg cttccgtgac ctttctcagc
     2461 atgaccaagt caccctgctt aaggctggca cctttgaggt gctgatggtg cgctttgctt
     2521 cgttgttcaa cgtgaaggac cagacagtga tgttcctaag ccgcaccacc tacagcctgc
     2581 aggagcttgg tgccatgggc atgggagacc tgctcagtgc catgttcgac ttcagcgaga
     2641 agctcaactc cctggcgctt accgaggagg agctgggcct cttcaccgcg gtggtgcttg
     2701 tctctgcaga ccgctcgggc atggagaatt ccgcttcggt ggagcagctc caggagacgc
     2761 tgctgcgggc tcttcgggct ctggtgctga agaaccggcc cttggagact tcccgcttca
     2821 ccaagctgct gctcaagctg ccggacctgc ggaccctgaa caacatgcat tccgagaagc
     2881 tgctgtcctt ccgggtggac gcccaggttt ctcaaggcag ttcaggtggc tcgagcggtg
     2941 tgagcggctg gcggctgttc aagaagatta gctaaggccg cgactctaga gtcgacctgc
     3001 aggcatgcaa gctgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     3061 caccgctgag caataactag cataacccct tggggcggcc gcttcgagca gacatgataa
     3121 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     3181 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     3241 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt
     3301 taagcaagta aaacctctac aaatgtggta aaatcgaatt ttaacaaaat attaacgctt
     3361 acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     3421 gcggatctgc gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac
     3481 cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3541 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3601 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3661 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3721 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3781 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3841 aaagcttaat taactgttga caattaatca tcggcatagt atatcggcat agtataatac
     3901 gacaaggtga ggaactaaac ccaggaggca gatcatgatt gaacaagatg gattgcacgc
     3961 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4021 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4081 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     4141 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4201 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4261 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4321 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4381 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4441 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4501 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4561 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4621 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4681 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     4741 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgatggcc gcaataaaat
     4801 atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag cgataaggat
     4861 cctctttgcg cttgcgtttt cccttgtcca gatagcccag tagctgacat tcatccgggg
     4921 tcagcaccgt ttctgcggac tggctttcta cccggtatca gctcactcaa aggcggtaat
     4981 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5041 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5101 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5161 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5221 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5281 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5341 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5401 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5461 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5521 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5581 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5641 gattacgcgc agaaaaaaag gatttcaaga agatcctttg atcttttcta cggggtctga
     5701 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5761 cttcacctag atccttttat agtccggaaa tacaggaacg cacgctggat ggcccttcgc
     5821 tgggatggtg aaaccatgaa aaatggcagc ttcagtggat taagtggggg taatgtggcc
     5881 tgtaccctct ggttgcatag gtattcatac ggttaaaatt tatcaggcgc gattgcggca
     5941 gtttttcggg tggtttgttg ccatttttac ctgtctgctg ccgtgatcgc gctgaacgcg
     6001 ttttagcggt gcgtacaatt aagggattat ggtaaatcca cttactgtct gccctcgtag
     6061 ccatcgagat aaaccgcagt actccggcca cgatgcgtcc ggcgtagagg atcgagatct
//
