LOCUS       sequence-217582-        5676 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 5676)
  AUTHORS   Pfleger CM, Lee E, Kirschner MW
  TITLE     Substrate recognition by the Cdc20 and Cdh1 components of the 
            anaphase-promoting complex.
  JOURNAL   Genes Dev. 2001 Sep 15. 15(18):2396-407.
  PUBMED    11562349
REFERENCE   2  (bases 1 to 5676)
  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: "Genes Dev. 
            2001 Sep 15. 15(18):2396-407."
COMMENT     Sequence Label: HA-cdh1
FEATURES             Location/Qualifiers
     source          1..5676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..984
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1019..1037
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1019..1036
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1110..1136
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1140..1166
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1770..1781)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2712..2846
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2736..2755)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2790..2809
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2959..2979)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2959..2977)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2998..3014)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2998..3014)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3011..3033)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3022..3038
                     /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(3046..3052,3053..3070,3071..3076))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3091..3112
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3229..3246)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3400..3988)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3480..3499)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4159..4950,4951..5019))
                     /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     4782..4801
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5020..5124)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5150..5605)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5287..5308)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5499..5518
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tcgaccgcca attcaatatg gcgtatatgg actcatgcca attcaatatg gtggatctgg
       61 acctgtgcca attcaatatg gcgtatatgg actcgtgcca attcaatatg gtggatctgg
      121 accccagcca attcaatatg gcggacttgg caccatgcca attcaatatg gcggacctgg
      181 cactgtgcca actggggagg ggtctacttg gcacggtgcc aagtttgagg aggggtcttg
      241 gccctgtgcc aagtccgcca tattgaattg gcatggtgcc aataatggcg gccatattgg
      301 ctatatgcca ggatcaatat ataggcaata tccaatatgg ccctatgcca atatggctat
      361 tggccaggtt caatactatg tattggccct atgccatata gtattccata tatgggtttt
      421 cctattgacg tagatagccc ctcccaatgg gcggtcccat ataccatata tggggcttcc
      481 taataccgcc catagccact cccccattga cgtcaatggt ctctatatat ggtctttcct
      541 attgacgtca tatgggcggt cctattgacg tatatggcgc ctcccccatt gacgtcaatt
      601 acggtaaatg gcccgcctgg ctcaatgccc attgacgtca ataggaccac ccaccattga
      661 cgtcaatggg atggctcatt gcccattcat atccgttctc acgcccccta ttgacgtcaa
      721 tgacggtaaa tggcccactt ggcagtacat caatatctat taatagtaac ttggcaagta
      781 cattactatt ggaagtacgc cagggtacat tggcagtact cccattgacg tcaatggcgg
      841 taaatggccc gcgatggctg ccaagtacat ccccattgac gtcaatgggg aggggcaatg
      901 acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg ggaggtctat
      961 ataagcaatg ctcgtttagg gaaccgccat tctgcctggg gacgtcggag caagcttgat
     1021 ttaggtgaca ctatagaata caagctactt gttctttttg caggatccca tcggggatgg
     1081 agctccaccg cggtggcggc cgcatctttt acccatacga tgttcctgac tatgcgggct
     1141 atccctatga cgtcccggac tatgcaggat ccgattcgaa ttcgatggac caggactatg
     1201 agcggcgcct gcttcgccag atcgtcatcc agaatgagaa cacgatgcca cgcgtcacag
     1261 agatgcggcg gaccctgacg cctgccagct ccccagtgtc ctcgcccagc aagcacggag
     1321 accgcttcat cccctccaga gccggagcca actggagcgt gaacttccac aggattaacg
     1381 agaatgagaa gtctcccagt cagaaccgga aagccaagga cgccacctca gacaacggca
     1441 aagacggcct ggcctactct gccctgctca agaatgagct gctgggtgcc ggcatcgaga
     1501 aggtgcagga cccgcagact gaggaccgca gactgcagcc ctccacgcct gagaagaagg
     1561 gtctgttcac gtattccctt agcaccaagc gctccagccc cgatgacggc aacgatgtgt
     1621 ctccctactc cctgtctccc gtcagcaaca agagccagaa gctgctccgg tccccccgga
     1681 aacccacccg caagatctcc aagatcccct tcaaggtgct ggacgcgccc gagctgcagg
     1741 acgacttcta cctcaatctg gtggactggt cgtccctcaa tgtgctcagc gtggggctag
     1801 gcacctgcgt gtacctgtgg agtgcctgta ccagccaggt gacgcggctc tgtgacctct
     1861 cagtggaagg ggactcagtg acctccgtgg gctggtctga gcgggggaac ctggtggcgg
     1921 tgggcacaca caagggcttc gtgcagatct gggacgcagc cgcagggaag aagctgtcca
     1981 tgttggaggg ccacacggca cgcgtcgggg cgctggcctg gaatgctgag cagctgtcgt
     2041 ccgggagccg cgaccgcatg atcctgcaga gggacatccg caccccgcca ctgcagtcgg
     2101 agcggcggct gcagggccac cggcaggagg tgtgcgggct caagtggtcc acagaccacc
     2161 agctcctcgc ctcggggggc aacgacaaca agctgctggt ctggaatcac tcgagcctga
     2221 gccccgtgca gcagtacacg gagcacctgg cggccgtgaa ggccatcgcc tggtccccac
     2281 atcagcacgg gctgctggcc tcggggggcg gcacagctga ccgctgtatc cgcttctgga
     2341 acacgctgac aggacaacca ctgcagtgta tcgacacggg ctcccaagtg tgcaatctgg
     2401 cctggtccaa gcacgccaac gagctggtga gcacgcacgg ctactcacag aaccagatcc
     2461 ttgtctggaa gtacccctcc ctgacccagg tggccaagct gaccgggcac tcctaccgcg
     2521 tgctgtacct ggcaatgtcc cctgatgggg aggccatcgt cactggtgct ggagacgaga
     2581 ccctgaggtt ctggaacgtc tttagcaaaa cccgttcgac aaaggagtct gtgtctgtgc
     2641 tcaacctctt caccaggatc cggtaatcta gcctctcgag cctctagaac tatagtgagt
     2701 cgtattacgt agatccagac atgataagat acattgatga gtttggacaa accacaacta
     2761 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     2821 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     2881 ttcaggggga ggtgtgggag gttttttaat tcgcggccgc ggcgccaatg cattgggccc
     2941 ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
     3001 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3061 gagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3121 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3181 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3241 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3301 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3361 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3421 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3481 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3541 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3601 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3661 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3721 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3781 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3841 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3901 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3961 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4021 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4081 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4141 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4201 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4261 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4321 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4381 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4441 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4501 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4561 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4621 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4681 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4741 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4801 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4861 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4921 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4981 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5041 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5101 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc
     5161 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     5221 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     5281 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     5341 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     5401 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     5461 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     5521 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     5581 cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa
     5641 gggcgatcgg tgcgggcctc ttcgctatta cgccag
//
