LOCUS       sequence-350309-        5068 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast shuttle plasmid that allows conversion between low copy, high 
            copy, and integrating vectors. Low copy, HIS3-selected..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5068)
  AUTHORS   Nickerson DP, Quinn MA, Milnes JM
  TITLE     Rapid conversion of replicating and integrating Saccharomyces 
            cerevisiae plasmid vectors via Cre recombinase.
  JOURNAL   G3 (Bethesda). 2021 Dec 8;11(12). pii: 6380430. doi: 
            10.1093/g3journal/jkab336.
  PUBMED    34599813
REFERENCE   2  (bases 1 to 5068)
  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: "G3 
            (Bethesda)."; date: "2021-12-8"; volume: "11(12). pii: 6380430. 
            doi"; pages: " 10.1093/g3journal/jkab336"
COMMENT     Sequence Label: pDN613
FEATURES             Location/Qualifiers
     source          1..5068
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    5..38
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    74..577
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     protein_bind    589..622
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     659..677
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(715..737)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     837..856
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1003..1190
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             1191..1853
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(2113..2568)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2250..2271)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2462..2481
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2545..2925)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     2694..2716
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2708..2725
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2709..2725
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2735..2754
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2735..2753
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2762..2869
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2778..2794
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2779..2795
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2829..2845)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2829..2845)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2882..2902)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2882..2900)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2921..2937)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2921..2937)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2934..2956)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2945..2961
                     /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(2969..2975,2976..2993,2994..2999))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3014..3035
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3152..3169)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3323..3911)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3403..3422)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4082..4873,4874..4942))
                     /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     4705..4724
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4943..5047)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acctataact tcgtataatg tatgctatac gaagttattg acgtccccga aaagtgccac
       61 ctgggtcctt ttcatcacgt gctataaaaa taattataat ttaaattttt taatataaat
      121 atataaatta aaaatagaaa gtaaaaaaag aaattaaaga aaaaatagtt tttgttttcc
      181 gaagatgtaa aagactctag ggggatcgcc aacaaatact accttttatc ttgctcttcc
      241 tgctctcagg tattaatgcc gaattgtttc atcttgtctg tgtagaagac cacacacgaa
      301 aatcctgtga ttttacattt tacttatcgt taatcgaatg tatatctatt taatctgctt
      361 ttcttgtcta ataaatatat atgtaaagta cgctttttgt tgaaattttt taaacctttg
      421 tttatttttt tttcttcatt ccgtaactct tctaccttct ttatttactt tctaaaatcc
      481 aaatacaaaa cataaaaata aataaacaca gagtaaattc ccaaattatt ccatcattaa
      541 aagatacgag gcgcgtgtaa gttacaggca agcgatccgt ccgacgtcat aacttcgtat
      601 aatgtatgct atacgaagtt attaagaaac cattattatc atgacattaa cctataaaaa
      661 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
      721 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
      781 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
      841 atcagagcag attgtactga gagtgcacca taaattcccg ttttaagagc ttggtgagcg
      901 ctaggagtca ctgccaggta tcgtttgaac acggcattag tcagggaagt cataacacag
      961 tcctttcccg caattttctt tttctattac tcttggcctc ctctagtaca ctctatattt
     1021 ttttatgcct cggtaatgat tttcattttt ttttttccac ctagcggatg actctttttt
     1081 tttcttagcg attggcatta tcacataatg aattatacat tatataaagt aatgtgattt
     1141 cttcgaagaa tatactaaaa aatgagcagg caagataaac gaaggcaaag atgacagagc
     1201 agaaagccct agtaaagcgt attacaaatg aaaccaagat tcagattgcg atctctttaa
     1261 agggtggtcc cctagcgata gagcactcga tcttcccaga aaaagaggca gaagcagtag
     1321 cagaacaggc cacacaatcg caagtgatta acgtccacac aggtataggg tttctggacc
     1381 atatgataca tgctctggcc aagcattccg gctggtcgct aatcgttgag tgcattggtg
     1441 acttacacat agacgaccat cacaccactg aagactgcgg gattgctctc ggtcaagctt
     1501 ttaaagaggc cctaggggcc gtgcgtggag taaaaaggtt tggatcagga tttgcgcctt
     1561 tggatgaggc actttccaga gcggtggtag atctttcgaa caggccgtac gcagttgtcg
     1621 aacttggttt gcaaagggag aaagtaggag atctctcttg cgagatgatc ccgcattttc
     1681 ttgaaagctt tgcagaggct agcagaatta ccctccacgt tgattgtctg cgaggcaaga
     1741 atgatcatca ccgtagtgag agtgcgttca aggctcttgc ggttgccata agagaagcca
     1801 cctcgcccaa tggtaccaac gatgttccct ccaccaaagg tgttcttatg tagtgacacc
     1861 gattatttaa agctgcagca tacgatatat atacatgtgt atatatgtat acctatgaat
     1921 gtcagtaagt atgtatacga acagtatgat actgaagatg acaaggtaat gcatcattct
     1981 atacgtgtca ttctgaacga ggcgcgcttt ccttttttct ttttgctttt tctttttttt
     2041 tctcttgaac tcgacggatc tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     2101 taccgcatca ggaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     2161 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     2221 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     2281 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     2341 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     2401 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     2461 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     2521 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc attcgccatt
     2581 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2641 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     2701 acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat agggcgaatt
     2761 gggtaccggg ccccccctcg aggtcgacgg tatcgataag cttgatatcg aattcctgca
     2821 gcccggggga tccactagtt ctagagcggc cgccaccgcg gtggagctcc agcttttgtt
     2881 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     2941 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     3001 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3061 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3121 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3181 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3241 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3301 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3361 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3421 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3481 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3541 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3601 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3661 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3721 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3781 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3841 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3901 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3961 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4021 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4081 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4141 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4201 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4261 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4321 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4381 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4441 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4501 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4561 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4621 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4681 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4741 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4801 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4861 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4921 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     4981 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5041 ttccgcgcac atttccccga aaagtgcc
//
