LOCUS       sequence-375781-        5362 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of 14xPP7 loops in Saccharomyces cerevisiae. Selection 
            by KanMX (flanked by LoxP sites for marker removal).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5362)
  AUTHORS   Donovan BT, Huynh A, Ball DA, Patel HP, Poirier MG, Larson DR, 
            Ferguson ML, Lenstra TL
  TITLE     Live-cell imaging reveals the interplay between transcription 
            factors, nucleosomes, and bursting.
  JOURNAL   EMBO J. 2019 Jun 17;38(12). pii: embj.2018100809. doi: 
            10.15252/embj.2018100809. Epub 2019 May 17.
  PUBMED    31101674
REFERENCE   2  (bases 1 to 5362)
  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: "EMBO J."; 
            date: "2019-06-17"; volume: "38(12). pii: embj.2018100809. doi"; 
            pages: " 10.15252/embj.2018100809. Epub 2019 May 17"
COMMENT     Sequence Label: 14x PP7 with loxP-kanMX-loxP
FEATURES             Location/Qualifiers
     source          1..5362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(1..792,793..861))
                     /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     624..643
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(862..966)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(992..1447)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1129..1150)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1341..1360
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1574..1596
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1588..1605
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1589..1605
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1615..1634
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1615..1633
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2572..2605
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            2660..4016
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        2660..3003
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             3004..3813
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3071..3090)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3819..4016
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    4079..4112
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(4163..4183)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4163..4181)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4202..4218)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4202..4218)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4215..4237)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4226..4242
                     /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(4250..4256,4257..4274,4275..4280))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4295..4316
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4433..4450)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4604..5192)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4684..4703)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
       61 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
      121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
      181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
      241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
      301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
      361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
      421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
      481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
      541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
      601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
      661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
      721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
      781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
      841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
      901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
      961 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa
     1021 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     1081 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     1141 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     1201 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     1261 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     1321 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     1381 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     1441 ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     1501 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     1561 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac
     1621 gactcactat agggcgaatt ggagctccac cgtacaaagt ggagcgagga gatcctaagg
     1681 tacctaattg cctagaaagg agcagacgat atggcgtcgc tccctgcagg tcgactctag
     1741 aaaccagcag agcatatggg ctcgctggct gcagtattcc cgggttcatt agatcctaag
     1801 gtacctaatt gcctagaaag gagcagacga tatggcgtcg ctccctgcag gtcgactcta
     1861 gaaaccagca gagcatatgg gctcgctggc tgcagtattc ccgggttcat tagatcctaa
     1921 ggtacctaat tgcctagaaa ggagcagacg atatggcgtc gctccctgca ggtcgactct
     1981 agaaaccagc agagcatatg ggctcgctgg ctgcagtatt cccgggttca ttagatccta
     2041 aggtacctaa ttgcctagaa aggagcagac gatatggcgt cgctccctgc aggtcgactc
     2101 tagaaaccag cagagcatat gggctcgctg gctgcagtat tcccgggttc attagatcct
     2161 aaggtaccta attgcctaga aaggagcaga cgatatggcg tcgctccctg caggtcgact
     2221 ctagaaacca gcagagcata tgggctcgct ggctgcagta ttcccgggtt cattagatcc
     2281 taaggtacct aattgcctag aaaggagcag acgatatggc gtcgctccct gcaggtcgac
     2341 tctagaaacc agcagagcat atgggctcgc tggctgcagt attcccgggt tcattagatc
     2401 ctaaggtacc taattgccta gaaaggagca gacgatatgg cgtcgctccc tgcaggtcga
     2461 ctctagaaac cagcagagca tatgggctcg ctggctgcag tattcccggg ttcattagat
     2521 ctcgagaacg gggctgcagg aattgtacgc tgcaggtcga caacccttaa tataacttcg
     2581 tataatgtat gctatacgaa gttattaggt ctagagatct gtttagcttg cctcgtcccc
     2641 gccgggtcac ccggccagcg acatggaggc ccagaatacc ctccttgaca gtcttgacgt
     2701 gcgcagctca ggggcatgat gtgactgtcg cccgtacatt tagcccatac atccccatgt
     2761 ataatcattt gcatccatac attttgatgg ccgcacggcg cgaagcaaaa attacggctc
     2821 ctcgctgcag acctgcgagc agggaaacgc tcccctcaca gacgcgttga attgtcccca
     2881 cgccgcgccc ctgtagagaa atataaaagg ttaggatttg ccactgaggt tcttctttca
     2941 tatacttcct tttaaaatct tgctaggata cagttctcac atcacatccg aacataaaca
     3001 accatgggta aggaaaagac tcacgtttcg aggccgcgat taaattccaa catggatgct
     3061 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     3121 cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     3181 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     3241 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     3301 cccggcaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     3361 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     3421 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     3481 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     3541 atgcataagc ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     3601 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     3661 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     3721 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     3781 cagtttcatt tgatgctcga tgagtttttc taatcagtac tgacaataaa aagattcttg
     3841 ttttcaagaa cttgtcattt gtatagtttt tttatattgt agttgttcta ttttaatcaa
     3901 atgttagcgt gatttatatt ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa
     3961 gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct atactgctgt
     4021 cgattcgata ctaacgccgc catccagtgt cgaaaacgag ctctcgagaa cccttaatat
     4081 aacttcgtat aatgtatgct atacgaagtt attaggtgat atcagatcca ctagtggcct
     4141 atgcggtacc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
     4201 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4261 ccgggagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4321 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4381 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4441 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4501 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4561 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4621 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4681 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4741 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4801 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4861 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4921 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4981 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5041 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5101 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5161 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5221 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5281 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5341 atgagtaaac ttggtctgac ag
//
