LOCUS       sequence-456579-        5697 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 5697)
  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 5697)
  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-cdc20
FEATURES             Location/Qualifiers
     source          1..5697
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             126..152
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             156..182
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1749..1883
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1773..1792)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1827..1846
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1996..2016)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1996..2014)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2035..2051)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2035..2051)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2048..2070)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2059..2075
                     /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(2083..2089,2090..2107,2108..2113))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2128..2149
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2266..2283)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2437..3025)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2517..2536)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3196..3987,3988..4056))
                     /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     3819..3838
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4057..4161)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4187..4642)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4324..4345)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4536..4555
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4713..5697
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcgg ggatggagct ccaccgcggt ggcggccgca
      121 tcttttaccc atacgatgtt cctgactatg cgggctatcc ctatgacgtc ccggactatg
      181 caggatccga ttcgaattcg atggcacagt tcgcgttcga gagtgacctg cactcgctgc
      241 ttcagctgga tgcacccatc cccaatgcac cccctgcgcg ctggcagcgc aaagccaagg
      301 aagccgcagg cccggccccc tcacccatgc gggccgccaa ccgatcccac agcgccggca
      361 ggactccggg ccgaactcct ggcaaatcca gttccaaggt tcagaccact cctagcaaac
      421 ctggcggtga ccgctatatc ccccatcgca gtgctgccca gatggaggtg gccagcttcc
      481 tcctgagcaa ggagaaccag cctgaaaaca gccagacgcc caccaagaag gaacatcaga
      541 aagcctgggc tttgaacctg aacggttttg atgtagagga agccaagatc cttcggctca
      601 gtggaaaacc acaaaatgcg ccagagggtt accagaacag actgaaagta ctctacagcc
      661 aaaaggccac tcctggctcc agccggaaga cctgccgtta cattccttcc ctgccagacc
      721 gtatcctgga tgcgcctgaa atccgaaatg actattacct gaaccttgtg gattggagtt
      781 ctgggaatgt actggccgtg gcactggaca acagtgtgta cctgtggagt gcaagctctg
      841 gtgacatcct gcagcttttg caaatggagc agcctgggga atatatatcc tctgtggcct
      901 ggatcaaaga gggcaactac ttggctgtgg gcaccagcag tgctgaggtg cagctatggg
      961 atgtgcagca gcagaagcgg cttcgaaata tgaccagtca ctctgcccga gtgggctccc
     1021 taagctggaa cagctatatc ctgtccagtg gttcacgttc tggccacatc caccaccatg
     1081 atgttcgggt agcagaacac catgtggcca cactgagtgg ccacagccag gaagtgtgtg
     1141 ggctgcgctg ggccccagat ggacgacatt tggccagtgg tggtaatgat aacctggtca
     1201 atgtgtggcc tagtgctcct ggagagggtg gctgggttcc tctgcagaca ttcacccagc
     1261 atcaaggggc tgtcaaggcc gtagcatggt gtccctggca gtccaatgtc ctggcaacag
     1321 gagggggcac cagtgatcga cacattcgca tctggaatgt gtgctctggg gcctgtctga
     1381 gtgccgtgga tgcccattcc caggtgtgct ccatcctctg gtctccccat tacaaggagc
     1441 tcatctcagg ccatggcttt gcacagaacc agctagttat ttggaagtac ccaaccatgg
     1501 ccaaggtggc tgaactcaaa ggtcacacat cccgggtcct gagtctgacc atgagcccag
     1561 atggggccac agtggcatcc gcagcagcag atgagaccct gaggctatgg cgctgttttg
     1621 agttggaccc tgcgcggcgg cgggagcggg agaaggccag tgcagccaaa agcagcctca
     1681 tccaccaagg catccgctga tctagagccc tctcgagcct ctagaactat agtgagtcgt
     1741 attacgtaga tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     1801 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     1861 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     1921 agggggaggt gtgggaggtt ttttaattcg cggccgcggc gccaatgcat tgggcccggt
     1981 acccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     2041 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccgggag
     2101 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     2161 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2221 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2281 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2341 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2401 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2461 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2521 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2581 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2641 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2701 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2761 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2821 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2881 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2941 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3001 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3061 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3121 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3181 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3241 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3301 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3361 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3421 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3481 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3541 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3601 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3661 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3721 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3781 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3841 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3901 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3961 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4021 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4081 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4141 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
     4201 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     4261 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     4321 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     4381 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     4441 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     4501 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     4561 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     4621 aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg
     4681 cgatcggtgc gggcctcttc gctattacgc cagtcgaccg ccaattcaat atggcgtata
     4741 tggactcatg ccaattcaat atggtggatc tggacctgtg ccaattcaat atggcgtata
     4801 tggactcgtg ccaattcaat atggtggatc tggaccccag ccaattcaat atggcggact
     4861 tggcaccatg ccaattcaat atggcggacc tggcactgtg ccaactgggg aggggtctac
     4921 ttggcacggt gccaagtttg aggaggggtc ttggccctgt gccaagtccg ccatattgaa
     4981 ttggcatggt gccaataatg gcggccatat tggctatatg ccaggatcaa tatataggca
     5041 atatccaata tggccctatg ccaatatggc tattggccag gttcaatact atgtattggc
     5101 cctatgccat atagtattcc atatatgggt tttcctattg acgtagatag cccctcccaa
     5161 tgggcggtcc catataccat atatggggct tcctaatacc gcccatagcc actcccccat
     5221 tgacgtcaat ggtctctata tatggtcttt cctattgacg tcatatgggc ggtcctattg
     5281 acgtatatgg cgcctccccc attgacgtca attacggtaa atggcccgcc tggctcaatg
     5341 cccattgacg tcaataggac cacccaccat tgacgtcaat gggatggctc attgcccatt
     5401 catatccgtt ctcacgcccc ctattgacgt caatgacggt aaatggccca cttggcagta
     5461 catcaatatc tattaatagt aacttggcaa gtacattact attggaagta cgccagggta
     5521 cattggcagt actcccattg acgtcaatgg cggtaaatgg cccgcgatgg ctgccaagta
     5581 catccccatt gacgtcaatg gggaggggca atgacgcaaa tgggcgttcc attgacgtaa
     5641 atgggcggta ggcgtgccta atgggaggtc tatataagca atgctcgttt agggaac
//
