LOCUS       sequence-64039-e        5814 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 5814)
  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 5814)
  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: pDZ415 (24MS2SL loxP-Kan-loxP)
FEATURES             Location/Qualifiers
     source          1..5814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            45..1401
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        45..388
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             389..1198
                     /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(456..475)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1204..1401
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    1464..1497
                     /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(1548..1568)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1548..1566)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1587..1603)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1587..1603)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1600..1622)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1611..1627
                     /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(1635..1641,1642..1659,1660..1665))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1680..1701
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1818..1835)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1989..2577)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2069..2088)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2748..3539,3540..3608))
                     /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     3371..3390
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3609..3713)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3740..4195
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3827..3846)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4037..4058
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4321..4343
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4335..4352
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4336..4352
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4362..4381
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4362..4380
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     4413..4429
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4413..4429
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    5771..5804
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 gatctgttta gcttgcctcg tccccgccgg gtcacccggc cagcgacatg gaggcccaga
       61 ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt
      121 acatttagcc catacatccc catgtataat catttgcatc catacatttt gatggccgca
      181 cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga aacgctcccc
      241 tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata aaaggttagg
      301 atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta ggatacagtt
      361 ctcacatcac atccgaacat aaacaaccat gggtaaggaa aagactcacg tttcgaggcc
      421 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
      481 cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt
      541 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
      601 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
      661 tgcatggtta ctcaccactg cgatccccgg caaaacagca ttccaggtat tagaagaata
      721 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
      781 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
      841 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
      901 gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac cggattcagt
      961 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     1021 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     1081 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     1141 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     1201 agtactgaca ataaaaagat tcttgttttc aagaacttgt catttgtata gtttttttat
     1261 attgtagttg ttctatttta atcaaatgtt agcgtgattt atattttttt tcgcctcgac
     1321 atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt caatcgtatg
     1381 tgaatgctgg tcgctatact gctgtcgatt cgatactaac gccgccatcc agtgtcgaaa
     1441 acgagctctc gagaaccctt aatataactt cgtataatgt atgctatacg aagttattag
     1501 gtgatatcag atccactagt ggcctatgcg gtacccagct tttgttccct ttagtgaggg
     1561 ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     1621 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     1681 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     1741 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     1801 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1861 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1921 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     1981 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2041 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2101 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2161 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2221 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2281 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2341 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2401 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     2461 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2521 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2581 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2641 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2701 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     2761 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     2821 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     2881 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     2941 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3001 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3061 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3121 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3181 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3241 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3301 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3361 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3421 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3481 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3541 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3601 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3661 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     3721 ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3781 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3841 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3901 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3961 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4021 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4081 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     4141 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttccatt
     4201 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     4261 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     4321 cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg
     4381 gcgaattgga gctccaccgc ggtggcggcc gctctagaac tagtggatcc tacggtactt
     4441 attgccaaga aagcacgagc atcagccgtg cctccaggtc gaatcttcaa acgacgacga
     4501 tcacgcgtcg ctccagtatt ccagggttca tcagatccta cggtacttat tgccaagaaa
     4561 gcacgagcat cagccgtgcc tccaggtcga atcttcaaac gacgacgatc acgcgtcgct
     4621 ccagtattcc agggttcatc agatcctacg gtacttattg ccaagaaagc acgagcatca
     4681 gccgtgcctc caggtcgaat cttcaaacga cgacgatcac gcgtcgctcc agtattccag
     4741 ggttcatcag atcctacggt acttattgcc aagaaagcac gagcatcagc cgtgcctcca
     4801 ggtcgaatct tcaaacgacg acgatcacgc gtcgctccag tattccaggg ttcatcagat
     4861 cctacggtac ttattgccaa gaaagcacga gcatcagccg tgcctccagg tcgaatcttc
     4921 aaacgacgac gatcacgcgt cgctccagta ttccagggtt catcagatcc tacggtactt
     4981 attgccaaga aagcacgagc atcagccgtg cctccaggtc gaatcttcaa acgacgacga
     5041 tcacgcgtcg ctccagtatt ccagggttca tcagatccta cggtacttat tgccaagaaa
     5101 gcacgagcat cagccgtgcc tccaggtcga atcttcaaac gacgacgatc acgcgtcgct
     5161 ccagtattcc agggttcatc agatcctacg gtacttattg ccaagaaagc acgagcatca
     5221 gccgtgcctc caggtcgaat cttcaaacga cgacgatcac gcgtcgctcc agtattccag
     5281 ggttcatcag atcctacggt acttattgcc aagaaagcac gagcatcagc cgtgcctcca
     5341 ggtcgaatct tcaaacgacg acgatcacgc gtcgctccag tattccaggg ttcatcagat
     5401 cctacggtac ttattgccaa gaaagcacga gcatcagccg tgcctccagg tcgaatcttc
     5461 aaacgacgac gatcacgcgt cgctccagta ttccagggtt catcagatcc tacggtactt
     5521 attgccaaga aagcacgagc atcagccgtg cctccaggtc gaatcttcaa acgacgacga
     5581 tcacgcgtcg ctccagtatt ccagggttca tcagatccta cggtacttat tgccaagaaa
     5641 gcacgagcat cagccgtgcc tccaggtcga atcttcaaac gacgacgatc acgcgtcgct
     5701 ccagtattcc agggttcatc agatcccccg ggctgcagga attgtacgct gcaggtcgac
     5761 aacccttaat ataacttcgt ataatgtatg ctatacgaag ttattaggtc taga
//
