LOCUS       sequence-235852-        5109 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 5109)
  AUTHORS   Shi H, Rojas R, Bonifacino JS, Hurley JH
  TITLE     The retromer subunit Vps26 has an arrestin fold and binds Vps35 
            through its C-terminal domain.
  JOURNAL   Nat Struct Mol Biol. 2006 Jun . 13(6):540-8.
  PUBMED    16732284
REFERENCE   2  (bases 1 to 5109)
  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: "Nat Struct 
            Mol Biol. 2006 Jun . 13(6):540-8."
COMMENT     Sequence Label: pmr101A-hVPS26
FEATURES             Location/Qualifiers
     source          1..5109
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     106..125
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     209..228
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        209..227
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    228..252
                     /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)."
     RBS             267..289
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1286..1303
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1387..1405)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1401..1448
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2050..2865)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2050..2865)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2773..2792
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2857..2876
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(3326..3343)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3460..4005)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(3540..3559)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      complement(4584..5039)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4721..4742)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4933..4952
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
       61 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
      121 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
      181 gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg
      241 gataacaatt cccctctaga aataattttg tttaacttta agaaggagat ataccatggg
      301 tatgagtttt cttggaggct tttttggtcc aatttgtgag atcgatattg ttcttaatga
      361 tggggaaacc aggaaaatgg cagaaatgaa aactgaagat ggcaaagtag aaaaacacta
      421 tctcttctat gacggagaat ccgtttcagg aaaggtaaac ctagccttta agcaacctgg
      481 aaagaggcta gaacaccaag gaattagaat tgaatttgta ggtcaaattg aacttttcaa
      541 tgacaagagt aatactcatg aatttgtaaa cctagtgaaa gaactagcct tacctggaga
      601 actgactcag agcagaagtt atgattttga atttatgcaa gttgaaaagc catatgaatc
      661 ttacatcggt gccaatgtcc gcttgaggta ttttcttaaa gtgacaatag tgagaagact
      721 gacagatttg gtaaaagagt atgatcttat tgttcaccag cttgccacct atcctgatgt
      781 taacaactct attaagatgg aagtgggcat tgaagattgt ctacatatag aatttgaata
      841 taataaatca aagtatcatt taaaggatgt gattgttgga aaaatttact tcttattagt
      901 aagaataaaa atacaacata tggagttaca gctgatcaaa aaagagatca caggaattgg
      961 acccagtacc acaacagaaa cagaaacaat cgccaaatat gaaataatgg atggtgcacc
     1021 agtaaaaggt gaatcaattc caataaggct atttttagca ggatatgacc caactccaac
     1081 aatgagagat gtgaacaaaa aattttcagt aaggtacttt ttgaatttag tgcttgttga
     1141 tgaggaagac cggaggtact tcaaacagca ggagataatt ttatggagaa aagctcctga
     1201 aaaactgagg aaacagagaa caaactttca ccagcgattt gaatctccag aatcacaggc
     1261 atctgccgaa cagcctgaaa tgggactggt tccgcgtgga tcccatcatc atcatcattg
     1321 actcgagctc aagcttggat ccggctgcta acaaagcccg aaaggaagct gagttggctg
     1381 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     1441 gttttttgct gaaaggagga actatatccg gatatccaca ggacgggtgt ggtcgccatg
     1501 atggtgacca ccatttgccc catatcaccg tcggtactga tcccgtcgtc aataaaccga
     1561 accgctacac cctgagcatc aaactctttt atcagttgga tcatgtcggc ggtgtcgcgg
     1621 ccaagacggt cgagcttctt caccagaatg acatcacctt cctccacctt catcctcagc
     1681 aaatccagcc cttcccgatc tgttgaactg ccggatgcct tgtcggtaaa gatgcggtta
     1741 gcttttaccc ctgcatcttt gagcgctgag gtctgcctcg tgaagaaggt gttgctgact
     1801 cataccaggc ctgaatcgcc ccatcatcca gccagaaagt gagggagcca cggttgatga
     1861 gagctttgtt gtaggtggac cagttggtga ttttgaactt ttgctttgcc acggaacggt
     1921 ctgcgttgtc gggaagatgc gtgatctgat ccttcaactc agcaaaagtt cgatttattc
     1981 aacaaagccg ccgtcccgtc aagtcagcgt aatgctctgc cagtgttaca accaattaac
     2041 caattctgat tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
     2101 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     2161 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     2221 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     2281 agtgacgact gaatccggtg agaatggcaa gagcttatgc atttctttcc agacttgttc
     2341 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     2401 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac
     2461 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     2521 atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa
     2581 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     2641 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     2701 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     2761 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     2821 taatcgcggc cttgagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt
     2881 actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat cttgtgcaat
     2941 gtaacatcag agattttgag acacaacgtg gctttgttga ataaatcgaa cttttgctga
     3001 gttgaaggat cagatcacgc atcttcccga caacgcagac cgttccgtgg caaagcaaaa
     3061 gttcaaaatc accaactggt ccacctacaa caaagctctc atcaaccgtg gctccctcac
     3121 tttctggctg gatgatgggg cgattcaggc ctggtatgag tcagcaacac cttcttcacg
     3181 aggcagacct cagcgctagc ggagtgtata ctggcttact atgttggcac tgatgagggt
     3241 gtcagtgaag tgcttcatgt ggcaggagaa aaaaggctgc accggtgcgt cagcagaata
     3301 tgtgatacag gatatattcc gcttcctcgc tcactgactc gctacgctcg gtcgttcgac
     3361 tgcggcgagc ggaaatggct tacgaacggg gcggagattt cctggaagat gccaggaaga
     3421 tacttaacag ggaagtgaga gggccgcggc aaagccgttt ttccataggc tccgcccccc
     3481 tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga caggactata
     3541 aagataccag gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg
     3601 gtttaccggt gtcattccgc tgttatggcc gcgtttgtct cattccacgc ctgacactca
     3661 gttccgggta ggcagttcgc tccaagctgg actgtatgca cgaacccccc gttcagtccg
     3721 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggaaaga catgcaaaag
     3781 caccactggc agcagccact ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc
     3841 ggttaaggct aaactgaaag gacaagtttt ggtgactgcg ctcctccaag ccagttacct
     3901 cggttcaaag agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa ggcggttttt
     3961 tcgttttcag agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatt
     4021 aaggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4081 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4141 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4201 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4261 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4321 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4381 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4441 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg gttgaaggct
     4501 ctcaagggca tcggtcgacg ctctccctta tgcgactcct gcgaaaagcc ccaaaaacag
     4561 gaagattgta taagcaaata tttaaattgt aaacgtcaat attttgttaa aattcgcgtt
     4621 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     4681 taaatcaaaa gaatagcccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     4741 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     4801 cccactacgt gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     4861 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     4921 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     4981 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     5041 ctcaattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa
     5101 tggtgcatg
//
