LOCUS       sequence-493024-        7234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Acyclovir-regulated Cre.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7234)
  AUTHORS   Hou Q, Oleynikov M, Mei X, Dong L, Hagen T, Jaffrey SR
  TITLE     A portable poison exon for small-molecule control of mammalian gene 
            expression.
  JOURNAL   Nat Methods. 2025 Nov 3. doi: 10.1038/s41592-025-02860-7.
  PUBMED    41184552
REFERENCE   2  (bases 1 to 7234)
  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. 2025 Nov 3. doi: 10.1038/s41592-025-02860-7."
COMMENT     Sequence Label: pcDNA3.1(+) Cyclone-Cre C-DYK
FEATURES             Location/Qualifiers
     source          1..7234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1143..1208
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1255..1272)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1261..1485
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1531..1959
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1618..1637)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1828..1849
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1968..1988)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1973..2302
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2153..2288
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2215..2234
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2369..3163
                     /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(2423..2442)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3033..3052
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3337..3458
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3374..3393)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3428..3447
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3507..3523)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3507..3523)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3520..3542)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3531..3547
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3555..3561,3562..3579,3580..3585))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3600..3621
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3738..3755)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3909..4497)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3989..4008)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4668..5459,5460..5528))
                     /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     5291..5310
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5529..5633)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5708..5727)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5899..6278
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6279..6482
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6433..6453
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6527..6546
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6527..6545
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      6592..6601
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 cacttagcct gggggtaact aaactggtcg agcgatggat ttccgtctct ggtgtagctg
       61 atgatccgaa taactacctg ttttgccggg tcagaaaaaa tggtgttgcc gcgccatctg
      121 ccaccagcca gctatcaact cgcgccctgg aagggatttt tgaagcaact catcgattga
      181 tttacggcgc taaggatgac tctggtcaga gatacctggc ctggtctgga cacagtgccc
      241 gtgtcggagc cgcgcgagat atggtaagta atcactcagc atcttttcct gacaattttt
      301 ttgtagttat gtgactttgt tttgttgatc atattttgtt gaataaaata agtaaaatgt
      361 cttgtgaaac aaaatgcttt ttaacatcca tataaagcta tctatatata gctatctatg
      421 tctatatagc tatttttttt aacttccttt attttcctta cagggtttca gacaaaatca
      481 aaaagaagga aggtgctcac agccaagcaa ttaaggagta agtgtaataa ccgaagtagt
      541 ttatgggcta ccgaaataca cttacgccag cattatgaaa gtgaatctta cttttgtaaa
      601 actttatggt ttgtggaaaa caaatgtttt tgaacattta aaaagttcag atgttaaaaa
      661 gttgaaaggt taatgtaaaa caatcaatat taaagaattt tgatgccaaa actattagat
      721 aaaaggttaa tctacatccc tactagaatt ctcatactta actggttggt tatgtggaag
      781 aaacatactt tcacaataaa gagctttagg atatgatgcc attttatatc actagtaggc
      841 agaccagcag actttttttt attgtgatat gggataacct aggcatactg cactgtacac
      901 tctgacatat gaagtgctct agtcaagttt aactggtgtc cacagaggac atggtttaac
      961 tggaattcgt caagcctctg gttctaattt ctcatttgca ggcccgcgct ggagtttcaa
     1021 taccggagat catgcaagct ggtggctgga ccaatgtaaa tattgtcatg aactatatcc
     1081 gtaacctgga tagtgaaaca ggggcaatgg tgcgcctgct ggaagatggc gacctcgccg
     1141 cggactacaa agaccatgac ggtgattata aagatcatga catcgattac aaggatgacg
     1201 atgacaagta acggccgctc gagtctagag ggcccgttta aacccgctga tcagcctcga
     1261 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     1321 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     1381 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     1441 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     1501 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     1561 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     1621 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     1681 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     1741 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     1801 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     1861 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     1921 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     1981 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2041 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2101 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     2161 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     2221 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     2281 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     2341 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     2401 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     2461 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     2521 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     2581 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     2641 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     2701 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     2761 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     2821 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     2881 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     2941 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     3001 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     3061 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     3121 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     3181 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     3241 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     3301 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     3361 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3421 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     3481 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     3541 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     3601 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     3661 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     3721 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3781 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3841 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3901 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3961 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4021 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4081 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4141 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4201 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4261 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4321 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4381 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4441 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4501 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4561 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4621 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4681 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4741 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4801 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4861 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4921 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4981 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5041 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5101 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5161 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5221 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5281 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5341 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5401 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5461 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5521 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5581 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5641 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     5701 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     5761 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     5821 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
     5881 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     5941 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     6001 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     6061 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     6121 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     6181 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     6241 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     6301 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     6361 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     6421 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     6481 ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat acgactcact
     6541 atagggagac ccaagctggc tagcgtttaa acttaagctt ggtaccgagc tgccaccatg
     6601 gcgagcaatt tactgaccgt acaccaaaat ttgcctgcat taccggtcga tgcaacgagt
     6661 gatgaggttc gcaagaacct gatggacatg ttcagggatc gccaggcgtt ttctgagcat
     6721 acctggaaaa tgcttctgtc cgtttgccgg tcgtgggcgg catggtgcaa gttgaataac
     6781 cggaaatggt ttcccgcaga acctgaagat gttcgcgatt atcttctata tcttcaggcg
     6841 cgcggtctgg cagtaaaaac tatccagcaa catttgggcc agctaaacat gcttcatcgt
     6901 cggtccgggc tgccacgacc aagtgacagc aatgctgttt cactggttat gcggcggatc
     6961 cgaaaagaaa acgttgatgc cggtgaacgt gcaaaacagg ctctagcgtt cgaacgcact
     7021 gatttcgacc aggttcgttc actcatggaa aatagcgatc gctgccagga tatacgtaat
     7081 ctggcatttc tggggattgc ttataacacc ctgttacgta tagccgaaat tgccaggatc
     7141 agggttaaag atatctcacg tactgacggt gggagaatgt taatccatat tggcagaacg
     7201 aaaacgctgg ttagcaccgc aggtgtagag aagg
//
