LOCUS       sequence-9958-ex        3935 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 3935)
  AUTHORS   Rudin N, Haber JE
  TITLE     Efficient repair of HO-induced chromosomal breaks in Saccharomyces 
            cerevisiae by recombination between flanking homologous sequences.
  JOURNAL   Mol Cell Biol. 1988 Sep . 8(9):3918-28.
  PUBMED    3065627
REFERENCE   2  (bases 1 to 3935)
  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: "Mol Cell 
            Biol. 1988 Sep . 8(9):3918-28."
COMMENT     Sequence Label: pNR6
FEATURES             Location/Qualifiers
     source          1..3935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..222
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             223..1026
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     primer_bind     complement(1180..1196)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1180..1196)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1193..1215)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1204..1220
                     /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(1228..1234,1235..1252,1253..1258))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1273..1294
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1411..1428)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1582..2170)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1662..1681)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2341..3132,3133..3201))
                     /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     2964..2983
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3202..3306)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3374..3392
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3430..3452)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3552..3571
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3765..3787
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3779..3796
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3780..3796
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aagcttttca attcatcatt ttttttttat tctttttttt gatttcggtt tccttgaaat
       61 ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga
      121 ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc
      181 caactgcaca gaacaaaaac ctgcaggaaa cgaagataaa tcatgtcgaa agctacatat
      241 aaggaacgtg ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac
      301 gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag
      361 ttagttgaag cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact
      421 gatttttcca tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt
      481 ttactcttcg aagacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct
      541 gcgggtgtat acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc
      601 ccaggtattg ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc
      661 cttttgatgt tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag
      721 ggtactgttg acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga
      781 gacatgggtg gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta
      841 gatgacaagg gagacgcatt gggtcaacag tatagaaccg tggatgatgt ggtctctaca
      901 ggatctgaca ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta
      961 gagggtgaac gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa
     1021 aactaaaaaa ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca
     1081 atttaattat atcagttatt acccgggaat ctcggtcgta atgattttta taatgacgaa
     1141 aaaaaaaaaa ttggaaagaa aaagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     1201 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     1261 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     1321 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     1381 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     1441 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     1501 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     1561 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     1621 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1681 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     1741 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     1801 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     1861 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     1921 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     1981 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     2041 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2101 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2161 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2221 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2281 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     2341 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     2401 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     2461 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     2521 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     2581 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     2641 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     2701 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     2761 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     2821 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     2881 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     2941 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     3001 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     3061 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     3121 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     3181 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     3241 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     3301 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     3361 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     3421 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     3481 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     3541 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     3601 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     3661 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     3721 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     3781 taaaacgacg gccagtgaat tcccggggat ctaaataaat tcgttttcaa tgattaaaat
     3841 agcatagtcg ggtttttctt ttagtttcag ctttccgcaa cagtataatt ttataaaccc
     3901 tggttttggt tttgtagagt ggttgacctg cagcc
//
