LOCUS       sequence-247044-        5694 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes for the CRY2-HA and mCherry proteins expressed from the EF1a
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5694)
  AUTHORS   Kim JH, Rege M, Valeri J, Dunagin MC, Metzger A, Titus KR, Gilgenast
            TG, Gong W, Beagan JA, Raj A, Phillips-Cremins JE
  TITLE     LADL: light-activated dynamic looping for endogenous gene expression
            control.
  JOURNAL   Nat Methods. 2019 Jun 24. pii: 10.1038/s41592-019-0436-5. doi: 
            10.1038/s41592-019-0436-5.
  PUBMED    31235883
REFERENCE   2  (bases 1 to 5694)
  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"; journalName: "Nat 
            Methods. 2019 Jun 24. pii: 10.1038/s41592-019-0436-5. doi: 
            10.1038/s41592-019-0436-5."
COMMENT     Sequence Label: Empty Target Control
FEATURES             Location/Qualifiers
     source          1..5694
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     603..623
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             683..730
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             2231..2251
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2282..2308
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2330..2383
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2384..3091
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2526..2544)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(3121..3138)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3127..3238
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        3374..3478
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3479..3547,3548..4339)
                     /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     complement(3697..3716)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4510..5098
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4999..5018
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5190..5401
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          join(5402..5694,1..646)
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
ORIGIN
        1 gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc acactggtat
       61 ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc
      121 gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg
      181 gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct
      241 ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg
      301 gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag
      361 gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc
      421 gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag gttgggggga
      481 ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc
      541 ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat
      601 tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgaaagct
      661 tggtaccgga tccgccacca tgaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa
      721 aaagaaaaag aagatggaca aaaagacgat tgtgtggttt agacgtgacc ttcgcattga
      781 ggacaaccct gcgttggcag cggcagcgca tgaaggttcg gtattccctg tctttatctg
      841 gtgtcccgaa gaggaggggc aattctatcc cggtcgcgcg tcccgatggt ggatgaaaca
      901 gtcgctcgca catcttagcc agtcgttgaa ggcgctcggc tctgatctga cgctcatcaa
      961 aacacacaat acaatttctg ccattctgga ttgcattagg gtgactggtg ccaccaaagt
     1021 tgtgttcaat cacctttatg atcccgtaag cctggtcagg gatcatacag taaaagagaa
     1081 acttgtagag aggggaatca gcgtccaatc gtacaatggg gacttgttgt atgaaccgtg
     1141 ggaaatctat tgtgagaaag gcaaaccttt cacttccttc aattcgtact ggaagaaatg
     1201 cctcgacatg tccatcgagt cagtcatgct gccgccacct tggagactta tgcccattac
     1261 tgctgcggca gaggcgatct gggcttgcag catcgaagaa cttggtctgg aaaacgaagc
     1321 ggaaaaaccc agtaacgcgc tcctgacgcg agcgtggtcg ccaggctggt cgaatgctga
     1381 taagctcctt aatgagttta tcgagaaaca gttgatcgac tatgcgaaga atagcaagaa
     1441 ggtggtggga aattccactt ccctcctgtc gccgtatctc cactttgggg agattagcgt
     1501 gaggcatgtg ttccaatgtg cgcggatgaa gcaaatcatt tgggctcgcg acaaaaactc
     1561 tgaaggagag gagtctgccg atttgttttt gagaggaatc ggccttaggg agtattcacg
     1621 atacatctgc ttcaactttc cgtttacaca tgaacaatcc ctcctcagcc acctccgctt
     1681 ctttccgtgg gacgcggacg tggacaaatt caaggcctgg cggcaggggc gcacgggata
     1741 cccgttggtg gatgcgggta tgagagagct gtgggcaacg ggttggatgc acaaccgtat
     1801 tcgagtcatc gttagtagct ttgccgttaa gttcctcttg ttgccctgga aatgggggat
     1861 gaaatacttt tgggacacgc ttctggatgc ggacttggag tgcgatattc ttgggtggca
     1921 gtacatttcg ggttccatcc ctgacgggca tgaacttgac cgattggata accctgcact
     1981 gcaaggagca aagtacgacc cggagggaga atacatcagg cagtggctcc cagagctggc
     2041 ccgtctgccc accgagtgga ttcaccaccc gtgggatgct cccctgaccg tccttaaagc
     2101 ctcaggggtc gaattgggga cgaattacgc gaaacccatc gtcgatattg acaccgcacg
     2161 agaattgctg gcgaaagcca tctcaagaac acgcgaagcg cagattatga tcggagcagc
     2221 gccggctagc cccaagaaga agagaaaggt ggaggccagc ggttccggac gggctgtata
     2281 ctacccgtac gacgttccgg actacgctct gtttaaactg ggcagtggag agggcagagg
     2341 aagtctgcta acatgcggtg acgtcgagga gaatcctggc ccagtgagca agggcgagga
     2401 ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt
     2461 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     2521 gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc
     2581 ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta
     2641 cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     2701 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt
     2761 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga aaaccatggg
     2821 ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa
     2881 gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa
     2941 ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac
     3001 ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc
     3061 caccggcggc atggacgagc tgtacaagta agaattcaat tcctagagct cgctgatcag
     3121 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     3181 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     3241 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     3301 aggattggga agagaatagc aggcatgatg aggagcgtct tagacgtcag gtggcacttt
     3361 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3421 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3481 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3541 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3601 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3661 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     3721 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     3781 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     3841 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     3901 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     3961 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4021 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4081 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4141 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtggaagccg
     4201 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4261 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4321 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4381 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4441 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4501 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4561 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4621 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4681 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4741 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4801 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4861 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4921 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4981 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5041 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5101 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5161 actacgtaaa tactagtgtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga
     5221 gaagttgggg ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa
     5281 ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta
     5341 tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca
     5401 ggtaagtgcc gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt
     5461 gccttgaatt acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg
     5521 aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt
     5581 tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg
     5641 tctcgctgct ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgc
//
