LOCUS       sequence-252816-        3270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 4 module-  CLN2-PEST (Degron).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3270)
  AUTHORS   Goulev Y, Matifas A, Heyer V, Reina-San-Martin B, Charvin G
  TITLE     COSPLAY: An expandable toolbox for combinatorial and swift 
            generation of expression plasmids in yeast.
  JOURNAL   PLoS One. 2019 Aug 28;14(8):e0220694. doi: 
            10.1371/journal.pone.0220694. eCollection 2019.
  PUBMED    31461461
REFERENCE   2  (bases 1 to 3270)
  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: "PLoS One.";
            date: "2019-08-28"; volume: "14(8):e0220694. doi"; pages: " 
            10.1371/journal.pone.0220694. eCollection 2019"
COMMENT     Sequence Label: pMV5
FEATURES             Location/Qualifiers
     source          1..3270
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     173..195
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     187..204
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     188..204
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(858..874)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(858..874)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(871..893)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    882..898
                     /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(906..912,913..930,931..936))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..972
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1089..1106)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1260..1848)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1340..1359)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2019..2810,2811..2879))
                     /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     2642..2661
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2880..2984)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3052..3070
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3108..3130)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3230..3249
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
       61 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      121 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      181 cgacgttgta aaacgacggc cagtgaattc gagctcggta cctcgcgaat gcatctagat
      241 atcgggatcc cggtctcgcg cagcatccaa cttgaacatt tcgagaaagc ttaccatatc
      301 aaccccatca tgctctttcg aaaattcaaa tagcacatcc attccttcgc ccgcttcctc
      361 atctcaaagc cacactccaa tgagaaacat gagctcactc tctgataaca gcgttttcag
      421 ccggaatatg gaacaatcat caccaatcac tccaagtatg taccaatttg gtcagcagca
      481 gtcaaacagt atatgtggta gcaccgttag tgtgaatagt ctggtgaata caaataacaa
      541 acaaaggatc tacgaacaaa tcacgggtcc taacagcaat aacgcaacca atgattatat
      601 tgatttgcta aacctaaatg agtctaacaa ggaaaaccaa aatcccgcaa cggcgcatta
      661 cctcaatggg ggcccaccca agacaagctt cattaaccat ggaatgttcc cctcgccaac
      721 tgggaccata aatagcggta aatctagcag tgcctcatct ttaatttctt ttggtatggg
      781 caatacccaa gtaatatagc aggagagacc gggcccgtcg actgcagagg cctgcatgca
      841 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
      901 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
      961 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     1021 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     1081 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     1141 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     1201 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     1261 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     1321 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     1381 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     1441 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     1501 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     1561 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     1621 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     1681 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     1741 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     1801 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     1861 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     1921 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     1981 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     2041 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     2101 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     2161 gatccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     2221 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     2281 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     2341 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     2401 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     2461 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     2521 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     2581 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     2641 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     2701 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     2761 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     2821 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     2881 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     2941 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     3001 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     3061 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     3121 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     3181 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     3241 cagattgtac tgagagtgca ccatatgcgg
//
