LOCUS       sequence-478479-        6146 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  ExpressHiBiT-NR1D1 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 6146)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6146)
  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            "Unpublished"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-NR1D1 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6146
                     /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(3089..3107)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3144..3265
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3155..3174)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3209..3228
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3510..3867
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3510..3701
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3533..3553)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3718..3853
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3882..3929
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3961..4755
                     /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(4015..4034)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4625..4644
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4819..4867
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5106..5694)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5186..5205)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5810..6093
                     /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 ggctagcgct caccatggtg agcggctggc
     1081 ggctgttcaa gaagattagc gggagctccg gtggctcgag cggtgcgatc gccatgacga
     1141 ccctggactc caacaacaac acaggtggcg tcatcaccta cattggctcc agtggctcct
     1201 ccccaagccg caccagccct gaatccctct atagtgacaa ctccaatggc agcttccagt
     1261 ccctgaccca aggctgtccc acctacttcc caccatcccc cactggctcc ctcacccaag
     1321 acccggctcg ctcctttggg agcattccac ccagcctgag tgatgacggc tccccttctt
     1381 cctcatcttc ctcgtcgtca tcctcctcct ccttctataa tgggagcccc cctgggagtc
     1441 tacaagtggc catggaggac agcagccgag tgtcccccag caagagcacc agcaacatca
     1501 ccaagctgaa tggcatggtg ttactgtgta aagtgtgtgg ggacgttgcc tcgggcttcc
     1561 actacggtgt gcacgcctgc gagggctgca agggcttttt ccgtcggagc atccagcaga
     1621 acatccagta caaaaggtgt ctgaagaatg agaattgctc catcgtccgc atcaatcgca
     1681 accgctgcca gcaatgtcgc ttcaagaagt gtctctctgt gggcatgtct cgagacgctg
     1741 tgcgttttgg gcgcatcccc aaacgagaga agcagcggat gcttgctgag atgcagagtg
     1801 ccatgaacct ggccaacaac cagttgagca gccagtgccc gctggagact tcacccaccc
     1861 agcaccccac cccaggcccc atgggcccct cgccaccccc tgctccggtc ccctcacccc
     1921 tggtgggctt ctcccagttt ccacaacagc tgacgcctcc cagatcccca agccctgagc
     1981 ccacagtgga ggatgtgata tcccaggtgg cccgggccca tcgagagatc ttcacctacg
     2041 cccatgacaa gctgggcagc tcacctggca acttcaatgc caaccatgca tcaggtagcc
     2101 ctccagccac caccccacat cgctgggaaa atcagggctg cccacctgcc cccaatgaca
     2161 acaacacctt ggctgcccag cgtcataacg aggccctaaa tggtctgcgc caggctccct
     2221 cctcctaccc tcccacctgg cctcctggcc ctgcacacca cagctgccac cagtccaaca
     2281 gcaacgggca ccgtctatgc cccacccacg tgtatgcagc cccagaaggc aaggcacctg
     2341 ccaacagtcc ccggcagggc aactcaaaga atgttctgct ggcatgtcct atgaacatgt
     2401 acccgcatgg acgcagtggg cgaacggtgc aggagatctg ggaggatttc tccatgagct
     2461 tcacgcccgc tgtgcgggag gtggtagagt ttgccaaaca catcccgggc ttccgtgacc
     2521 tttctcagca tgaccaagtc accctgctta aggctggcac ctttgaggtg ctgatggtgc
     2581 gctttgcttc gttgttcaac gtgaaggacc agacagtgat gttcctaagc cgcaccacct
     2641 acagcctgca ggagcttggt gccatgggca tgggagacct gctcagtgcc atgttcgact
     2701 tcagcgagaa gctcaactcc ctggcgctta ccgaggagga gctgggcctc ttcaccgcgg
     2761 tggtgcttgt ctctgcagac cgctcgggca tggagaattc cgcttcggtg gagcagctcc
     2821 aggagacgct gctgcgggct cttcgggctc tggtgctgaa gaaccggccc ttggagactt
     2881 cccgcttcac caagctgctg ctcaagctgc cggacctgcg gaccctgaac aacatgcatt
     2941 ccgagaagct gctgtccttc cgggtggacg cccaggttta aacgaattcg ggctcggtac
     3001 ccggggatcc tctagagtcg acctgcaggc atgcaagctg atccggctgc taacaaagcc
     3061 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     3121 gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     3181 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3241 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3301 tcagggggag atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat
     3361 cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca
     3421 tctgtgcggt atttcacacc gcatacgcgg atctgcgcag caccatggcc tgaaataacc
     3481 tctgaaagag gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg
     3541 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3601 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3661 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3721 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3781 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3841 ttttttggag gcctaggctt ttgcaaaaag cttaattaac tgttgacaat taatcatcgg
     3901 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaacccag gaggcagatc
     3961 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4021 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4081 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4141 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4201 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4261 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4321 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4381 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4441 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     4501 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4561 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4621 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4681 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4741 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4801 tgccatcacg atggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt
     4861 ttgtgtgaat cgatagcgat aaggatcctc tttgcgcttg cgttttccct tgtccagata
     4921 gcccagtagc tgacattcat ccggggtcag caccgtttct gcggactggc tttctacccg
     4981 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5041 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5101 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5161 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5221 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5281 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5341 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5401 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5461 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5521 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     5581 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5641 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat
     5701 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5761 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttatagtc cggaaataca
     5821 ggaacgcacg ctggatggcc cttcgctggg atggtgaaac catgaaaaat ggcagcttca
     5881 gtggattaag tgggggtaat gtggcctgta ccctctggtt gcataggtat tcatacggtt
     5941 aaaatttatc aggcgcgatt gcggcagttt ttcgggtggt ttgttgccat ttttacctgt
     6001 ctgctgccgt gatcgcgctg aacgcgtttt agcggtgcgt acaattaagg gattatggta
     6061 aatccactta ctgtctgccc tcgtagccat cgagataaac cgcagtactc cggccacgat
     6121 gcgtccggcg tagaggatcg agatct
//
