LOCUS       sequence-64090-e        5970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5970)
  AUTHORS   Hocine S, Raymond P, Zenklusen D, Chao JA, Singer RH
  TITLE     Single-molecule analysis of gene expression using two-color RNA 
            labeling in live yeast.
  JOURNAL   Nat Methods. 2013 Feb;10(2):119-21. doi: 10.1038/nmeth.2305. Epub 
            2012 Dec 23.
  PUBMED    23263691
REFERENCE   2  (bases 1 to 5970)
  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: 
            "10.1038/nmeth.2305"; journalName: "Nat Methods"; date: "2013-02"; 
            volume: "10"; issue: "2"; pages: "119-21"
COMMENT     Sequence Label: pDZ416 (24PP7SL loxP-Kan-loxP)
FEATURES             Location/Qualifiers
     source          1..5970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1503..1536
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            1591..2947
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1591..1934
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1935..2744
                     /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(2002..2021)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2750..2947
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    3010..3043
                     /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(3094..3114)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3094..3112)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3133..3149)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3133..3149)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3146..3168)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3157..3173
                     /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(3181..3187,3188..3205,3206..3211))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3226..3247
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3364..3381)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3535..4123)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3615..3634)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4294..5085,5086..5154))
                     /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     4917..4936
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5155..5259)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5286..5741
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5373..5392)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5583..5604
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5867..5889
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5881..5898
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5882..5898
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5908..5927
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5908..5926
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     join(5959..5970,1..5)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(5959..5970,1..5)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 ggatcctaag gtacctaatt gcctagaaag gagcagacga tatggcgtcg ctccctgcag
       61 gtcgactcta gaaaccagca gagcatatgg gctcgctggc tgcagtattc ccgggttcat
      121 tagatcctaa ggtacctaat tgcctagaaa ggagcagacg atatggcgtc gctccctgca
      181 ggtcgactct agaaaccagc agagcatatg ggctcgctgg ctgcagtatt cccgggttca
      241 ttagatccta aggtacctaa ttgcctagaa aggagcagac gatatggcgt cgctccctgc
      301 aggtcgactc tagaaaccag cagagcatat gggctcgctg gctgcagtat tcccgggttc
      361 attagatcct aaggtaccta attgcctaga aaggagcaga cgatatggcg tcgctccctg
      421 caggtcgact ctagaaacca gcagagcata tgggctcgct ggctgcagta ttcccgggtt
      481 cattagatcc taaggtacct aattgcctag aaaggagcag acgatatggc gtcgctccct
      541 gcaggtcgac tctagaaacc agcagagcat atgggctcgc tggctgcagt attcccgggt
      601 tcattagatc ctaaggtacc taattgccta gaaaggagca gacgatatgg cgtcgctccc
      661 tgcaggtcga ctctagaaac cagcagagca tatgggctcg ctggctgcag tattcccggg
      721 ttcattagat cctaaggtac ctaattgcct agaaaggagc agacgatatg gcgtcgctcc
      781 ctgcaggtcg actctagaaa ccagcagagc atatgggctc gctggctgca gtattcccgg
      841 gttcattaga tcctaaggta cctaattgcc tagaaaggag cagacgatat ggcgtcgctc
      901 cctgcaggtc gactctagaa accagcagag catatgggct cgctggctgc agtattcccg
      961 ggttcattag atcctaaggt acctaattgc ctagaaagga gcagacgata tggcgtcgct
     1021 ccctgcaggt cgactctaga aaccagcaga gcatatgggc tcgctggctg cagtattccc
     1081 gggttcatta gatcctaagg tacctaattg cctagaaagg agcagacgat atggcgtcgc
     1141 tccctgcagg tcgactctag aaaccagcag agcatatggg ctcgctggct gcagtattcc
     1201 cgggttcatt agatcctaag gtacctaatt gcctagaaag gagcagacga tatggcgtcg
     1261 ctccctgcag gtcgactcta gaaaccagca gagcatatgg gctcgctggc tgcagtattc
     1321 ccgggttcat tagatcctaa ggtacctaat tgcctagaaa ggagcagacg atatggcgtc
     1381 gctccctgca ggtcgactct agaaaccagc agagcatatg ggctcgctgg ctgcagtatt
     1441 cccgggttca ttagatcccc cgggctgcag gaattgtacg ctgcaggtcg acaaccctta
     1501 atataacttc gtataatgta tgctatacga agttattagg tctagagatc tgtttagctt
     1561 gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac cctccttgac
     1621 agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat ttagcccata
     1681 catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc gcgaagcaaa
     1741 aattacggct cctcgctgca gacctgcgag cagggaaacg ctcccctcac agacgcgttg
     1801 aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt gccactgagg
     1861 ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca catcacatcc
     1921 gaacataaac aaccatgggt aaggaaaaga ctcacgtttc gaggccgcga ttaaattcca
     1981 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     2041 cgacaatcta tcgattgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     2101 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     2161 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     2221 ccactgcgat ccccggcaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     2281 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     2341 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
     2401 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     2461 tctggaaaga aatgcataag cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     2521 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     2581 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     2641 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     2701 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagta ctgacaataa
     2761 aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg tagttgttct
     2821 attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca tctgcccaga
     2881 tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc
     2941 tatactgctg tcgattcgat actaacgccg ccatccagtg tcgaaaacga gctctcgaga
     3001 acccttaata taacttcgta taatgtatgc tatacgaagt tattaggtga tatcagatcc
     3061 actagtggcc tatgcggtac ccagcttttg ttccctttag tgagggttaa ttgcgcgctt
     3121 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     3181 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     3241 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     3301 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3361 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3421 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3481 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3541 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3601 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3661 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3721 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3781 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3841 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3901 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3961 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4021 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4081 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4141 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4201 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4261 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4321 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4381 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4441 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4501 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4561 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4621 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4681 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4741 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4801 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4861 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4921 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4981 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5041 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5101 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5161 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5221 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5281 acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5341 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     5401 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     5461 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     5521 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5581 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     5641 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5701 atttaacgcg aattttaaca aaatattaac gcttacaatt tccattcgcc attcaggctg
     5761 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     5821 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     5881 tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga attggagctc
     5941 caccgcggtg gcggccgctc tagaactagt
//
