LOCUS       sequence-229132-        5324 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This was used as a nuclear marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5324)
  AUTHORS   Huang N, Chelliah Y, Shan Y, Taylor CA, Yoo SH, Partch C, Green CB, 
            Zhang H, Takahashi JS
  TITLE     Crystal structure of the heterodimeric CLOCK:BMAL1 transcriptional 
            activator complex.
  JOURNAL   Science. 2012 Jul 13;337(6091):189-94. doi: 10.1126/science.1222804.
            Epub 2012 May 31.
  PUBMED    22653727
REFERENCE   2  (bases 1 to 5324)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1126/science.1222804"; journalName: "Science"; date: 
            "2012-07-13- 13"; volume: "337"; issue: "6091"; pages: "189-94"
COMMENT     Sequence Label: LV-H2B-mRFP1
FEATURES             Location/Qualifiers
     source          1..5324
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(42..61)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        233..612
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        613..816
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     767..787
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1377..2051
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             1377..2051
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(1507..1525)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1775..1794
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1969..1992
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    complement(2119..2152)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2208..2796
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2261..2281)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2679..2690)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2799..2815)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2800..2816)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             3325..3505
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(3567..4155)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3647..3666)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4326..5117,5118..5186))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4949..4968
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5187..5291)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
       61 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
      121 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
      181 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
      241 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      301 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      361 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      421 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      481 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      541 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      601 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      661 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      721 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      781 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc
      841 tagcgctacc ggactcagat ctcgagctcg gtacccgggg atcctctaga gtctagacac
      901 gagcttttat ttaagtgcgt tcattctcac tgctgttatt gttttctgac agcatgcctg
      961 aaccagctaa gtcagctcct gctccgaaga agggttccaa gaaggctgta accaaggcgc
     1021 agaagaagga tggcaagaag cgcaagcgca gtcgtaagga gagctactcc gtgtatgtgt
     1081 acaaggtgct aaaacaggtt caccccgata ctggcatctc atccaaggcc atgggcatca
     1141 tgaattcctt cgttaacgac atcttcgaac gcatcgcagg cgaggcttcc cgtctggccc
     1201 actacaacaa gcgctcgacc attacctcca gggagatcca gaccgccgtg cgtctgctgc
     1261 ttcccggaga gctggccaag cacgcagtgt ccgaaggtac caaggctgtc accaagtata
     1321 caagctccaa gctgcagtcg actctagact cgagggatcc agatctccag tcttccatgg
     1381 cctcctccga ggacgtcatc aaggagttca tgcgcttcaa ggtgcgcatg gagggctccg
     1441 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
     1501 agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg gacatcctgt
     1561 cccctcagtt ccagtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact
     1621 acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
     1681 gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg
     1741 tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg
     1801 gctgggaggc ctccaccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca
     1861 agatgaggct gaagctgaag gacggcggcc actacgacgc cgaggtcaag accacctaca
     1921 tggccaagaa gcccgtgcag ctgcccggcg cctacaagac cgacatcaag ctggacatca
     1981 cctcccacaa cgaggactac accatcgtgg aacagtacga gcgcgccgag ggccgccact
     2041 ccaccggcgc ctctcgagcg ggatccacta gtccagtgtg gtggaattaa ttctgcagat
     2101 aattcgtcga gggacctaat aacttcgtat agcatacatt atacgaagtt atacatgttt
     2161 aagggttccg gttccactag gtacaattcg atatcaagct tatcgataat caacctctgg
     2221 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     2281 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     2341 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     2401 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     2461 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     2521 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     2581 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
     2641 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     2701 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     2761 agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgatcga
     2821 gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt
     2881 gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct
     2941 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
     3001 ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac
     3061 cacacacaag gctacttccc tgattggcag aactacacac cagggccagg gatcagatat
     3121 ccactgacct ttggatggtg ctacaagcta gtaccagttg agcaagagaa ggtagaagaa
     3181 gccaatgaag gagagaacac ccgcttgtta caccctgtga gcctgcatgg gatggatgac
     3241 ccggagagag aagtattaga gtggaggttt gacagccgcc tagcatttca tcacatggcc
     3301 cgagagctgc atccggactg tactgggtct ctctggttag accagatctg agcctgggag
     3361 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     3421 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     3481 tagtcagtgt ggaaaatctc tagcagcatg tgagcaaaag gccagcaaaa ggccaggaac
     3541 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3601 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3661 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3721 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3781 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3841 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3901 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3961 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4021 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4081 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4141 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4201 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4261 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4321 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4381 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4441 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4501 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4561 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4621 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4681 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4741 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4801 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4861 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4921 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4981 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5041 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5101 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5161 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5221 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5281 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcga
//
