LOCUS       sequence-450341-        6032 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 6032)
  AUTHORS   Hierro A, Sun J, Rusnak AS, Kim J, Prag G, Emr SD, Hurley JH
  TITLE     Structure of the ESCRT-II endosomal trafficking complex.
  JOURNAL   Nature. 2004 Sep 9. 431(7005):221-5.
  PUBMED    15329733
REFERENCE   2  (bases 1 to 6032)
  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: "Nature. 
            2004 Sep 9. 431(7005):221-5."
COMMENT     Sequence Label: ESCRT-II
FEATURES             Location/Qualifiers
     source          1..6032
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     27..46
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        27..45
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             77..99
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     740..757
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     741..757
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             813..835
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             855..872
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             894..914
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     RBS             1669..1691
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3462..3480)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3476..3523
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3590..3681
                     /gene="bla"
                     /label=AmpR promoter
     terminator      4546..4585
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      4616..5204
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5105..5124
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5358..5375
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5390..5529
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5455..5474)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5615..5637
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5631..5822)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ggcaccatgc atcgatcccg cgaaattaat acgactcact atagggagac cacaacggtt
       61 tccctctaga aataattttg tttaacttta agaaggagat atacatatgt ctgcattacc
      121 tccagtatac tcattccctc cactatatac ccgtcaaccc aattctttaa caaggagaca
      181 acagataagt acgtggatag acatcatatc acaatattgc aagaccaaga aaatttggta
      241 catgtctgtt gacgggactg taatcaatga caacgaacta gatagtggaa gtacagataa
      301 tgacgattca aaaaaaatct cgaaaaatct ttttaataat gaggacatcc aaagatctgt
      361 atcacaggta tttattgatg aaatttggtc gcaaatgacc aaagaaggca aatgcttacc
      421 gattgaccaa agtggtagga gaagcagcaa taccaccact acgagatatt ttatattatg
      481 gaagagctta gacagttggg cttctttgat actacagtgg ttcgaagact ctggaaaact
      541 gaatcaagtc attacgttat acgaattgag tgaaggtgat gaaacagtca attgggagtt
      601 ccatagaatg cccgaaagct tgctctatta ttgtttgaag cccctatgcg atagaaatag
      661 ggccacaatg ctgaaagatg aaaatgataa agtaattgcc attaaggttg tttaaggatc
      721 cgggccctgt acaagatcct gtaaaacgac ggccagtgaa ttccccggga agcttcgcca
      781 gggttttccc agtcgagctc tctagaaata attttgttta actttaagaa ggagatatac
      841 atatgtcgta ctaccatcac catcaccatc acgattacga tatcccaacg accgaaaacc
      901 tgtattttca gggcgccatg aaacagtttg gactggcagc ctttgacgag ttgaaggatg
      961 gaaagtacaa tgacgtaaac aaaaccatac ttgaaaagca atctgtcgaa ctaagggacc
     1021 aattaatggt attccaagaa cgactggtag aatttgccaa gaaacacaat agcgagcttc
     1081 aagcttcacc tgaattccgc tcaaaattca tgcatatgtg ctcatctatt gggattgatc
     1141 ccctatcttt atttgatcga gataagcact tatttacagt gaatgatttt tattatgaag
     1201 tatgtcttaa agttatagaa atttgtcggc aaactaaaga tatgaatgga ggcgtcattt
     1261 cttttcaaga gttggagaag gtgcattttc gtaagctgaa tgttggactc gatgatttgg
     1321 aaaaatctat cgacatgcta aagagcctag agtgttttga gatatttcaa attcgcggca
     1381 aaaagttttt acgaagtgtt ccaaatgagt tgacatcaga tcaaacaaag attttggaaa
     1441 tatgctcaat tctgggctac tcaagtatct ctttattaaa ggcaaacttg ggatgggagg
     1501 ctgttaggag taaatctgct cttgatgaaa tggtcgctaa cggtttattg tggattgatt
     1561 atcaaggtgg tgcagaggcg ttgtattggg atccctcttg gattacgagg caactgtgag
     1621 gtaccagcgg ataacaattt cacatccgga gagctctcta gaaataattt tgtttaactt
     1681 taagaaggag atatacatat ggagtactgg cattatgtgg aaactacgtc atcgggccaa
     1741 cctcttcttc gagaaggtga aaaagatatc tttatcgatc agtccgtagg tctttaccat
     1801 ggcaaatcta agattctaca gcgacagagg ggcagaatct ttttaacgtc acaaagaatt
     1861 atctacatcg acgatgcaaa gcccactcag aactcgcttg ggttggaatt ggacgatcta
     1921 gcgtatgtaa actattcatc tggcttcttg accagatcac ctcgtctgat cctgtttttc
     1981 aaggacccct ccagtaagga tgaacttggt aaaagtgcgg aaacagcaag tgccgacgtt
     2041 gtttcgacgt gggtctgccc catctgtatg gtttcgaacg aaacgcaggg ggaattcact
     2101 aaagacactc tacctacacc catttgtatc aattgcggtg tgccagcgga ctatgagctc
     2161 accaagtcgt ctataaattg ttctaatgca atcgacccga acgccaaccc acagaatcag
     2221 ttcggagtga actctgagaa catttgcccc gcctgtactt ttgctaatca tcctcaaata
     2281 ggaaattgcg agatttgcgg ccacagattg cctaatgcat ccaaagtacg atcaaagctg
     2341 aacaggctaa actttcatga ttctagggtc catattgagc tagaaaaaaa ttcactggct
     2401 agaaacaaga gctcgcactc ggcattatcg tcctcgtcct cgacaggatc atcgacagaa
     2461 ttcgtgcaat tgagtttccg caaatctgat ggcgtattgt tttcacaagc tacagagagg
     2521 gctttggaga atatactcac tgaaaagaat aagcatattt tcaatcagaa tgtggtttct
     2581 gtaaatggtg tagacatgag gaaaggggct agttctcacg aatacaacaa tgaggtgcca
     2641 tttattgaga ctaaattgag cagaattggc atatcaagct tggaaaaatc tagagaaaac
     2701 cagcttttga acaatgacat tctctttaac aatgcgttga ccgacctaaa caagctaatg
     2761 tcgctggcta ccagcatcga aagactatac aaaaatagca acataacgat gaaaaccaag
     2821 acattgaact tacaggatga atcaaccgtg aatgaaccga aaacgagaag accgctattg
     2881 atacttgata gagagaagtt cctaaataaa gagctgttct tggatgagat tgcgagagaa
     2941 atttacgagt tcacgttgtc cgagtttaaa gatttgaaca gtgataccaa ctatatgatc
     3001 ataactctgg tagatctata tgcaatgtac aacaaatcta tgcgtatagg tacaggcctg
     3061 atatctccca tggaaatgag agaagcatgc gaaagattcg agcatttagg tctaaatgaa
     3121 ttgaagttag tgaaagttaa caaaagaata ttatgtgtaa cgagcgaaaa attcgacgtt
     3181 gtaaaagaaa agctagtaga tctgatcggt gataatcctg gttctgatct cctaagattg
     3241 acacagattt tgagttccaa taattcaaaa tcaaactgga cgctaggtat ccttatggag
     3301 gtattacaaa attgtgtcga tgaaggcgat ttactgatag acaagcaact gagtgggatt
     3361 tactactata aaaactctta ttggccctcg catatataaa cgcgtctcga gagatccggc
     3421 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     3481 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
     3541 atccgactag tagcccgcct aatgagcggg ctttttttta attcccctat ttgtttattt
     3601 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3661 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     3721 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctcgtgaa agtaaaagac
     3781 gcagaggacc aattgggggc acgagtggga tacatagaac tggacttgaa tagcggtaaa
     3841 atccttgaga gttttcgccc tgaagagcgt tttccaatga tgagcacttt caaagttctg
     3901 ctatgtggag cagtattatc ccgtgtagat gcggggcaag agcaactcgg acgacgaata
     3961 cactattcgc agaatgactt ggttgaatac tccccagtga cagaaaagca ccttacggac
     4021 ggaatgacgg taagagaatt atgtagtgcc gccataacga tgagtgataa cactgcggcg
     4081 aacttacttc tgacaaccat cggtggaccg aaggaattaa ccgctttttt gcacaatatg
     4141 ggagaccatg taactcgcct tgaccgttgg gaaccagaac tgaatgaagc cataccaaac
     4201 gacgagcgag acaccacaat gcctgcggca atggcaacaa cattacgcaa actattaact
     4261 ggcgaactac ttactctggc ttcacggcaa caattaatag actggcttga agcggataaa
     4321 gttgcaggac cactactgcg ttcggcactt cctgctggct ggtttattgc tgataaatct
     4381 ggggcaggag agcgtggttc acggggtatc attgccgcac ttggaccaga tggtaagcct
     4441 tcccgtatcg tagttatcta cacgacgggt agtcaggcaa ctatggacga acgaaataga
     4501 cagattgctg aaataggggc ttcactgatt aagcattggt aaaccgatac aattaaaggc
     4561 tccttttgga gccttttttt ttggacggac cggtagaaaa gatcaaagga tcttcttgag
     4621 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4681 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4741 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     4801 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4861 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4921 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4981 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5041 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5101 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5161 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5221 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     5281 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     5341 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     5401 attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat
     5461 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     5521 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     5581 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     5641 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     5701 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     5761 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     5821 actgatgcct ccgtgtaagg gggatttctg ttcatggggt aatgataccg atgaaacgag
     5881 agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg
     5941 agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat
     6001 gccagcgctt cgttaataca gatgtaggtg tt
//
