LOCUS       sequence-200252-        7452 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express His Tag Ate1-4 1A7B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7452)
  TITLE     Varshavsky lab plasmids
REFERENCE   2  (bases 1 to 7452)
  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"
COMMENT     Sequence Label: pCB410 (His-Ub-Ate1-4 1A7B)
FEATURES             Location/Qualifiers
     source          1..7452
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     969..988
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1008..1085)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1291..1310
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1394..1413
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1394..1412
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1413..1437
                     /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             1452..1474
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1493..1522
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1532..1549
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1553..1780
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     primer_bind     complement(3373..3391)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3387..3434
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3637..3655)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3723..3827
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3828..3896,3897..4688)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4046..4065)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4859..5447
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5348..5367
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5601..5618
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5633..5772
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5698..5717)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5858..5880
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5874..6065)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7377..7452,1..1007))
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga
       61 ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc
      121 agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg
      181 gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg
      241 gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg
      301 ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc
      361 cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc
      421 agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat
      481 gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg
      541 atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc
      601 acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc
      661 gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac
      721 accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac
      781 ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc
      841 agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt
      901 tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa
      961 gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg
     1021 aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg
     1081 gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag
     1141 taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc
     1201 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
     1261 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac
     1321 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc
     1381 gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa caattcccct
     1441 ctagaaataa ttttgtttaa ctttaagaag gagatatacc atgggatctt ctcaccacca
     1501 ccaccatcat catcatcatc acagcagcgg cctggtgccg cgcggcagcc atatgcagat
     1561 ctttgtgaag accctcactg gcaaaaccat cacccttgag gtcgagccca gtgacaccat
     1621 tgagaatgtc aaagccaaaa ttcaagacaa ggagggtatc ccacctgacc agcagcgtct
     1681 gatatttgcc ggcaaacagc tggaggatgg ccgcactctc tcagactaca acatccagaa
     1741 agagtccacc ctgcacctgg tgttgcgcct ccgcggtggt atggcctcgg tggtggaata
     1801 caagggcctg aaggccggct actactgcgg ctactgcgag tccagggagg gcaagacgtc
     1861 gtgtggcatg tgggcacatt ccatgacagt gcaggattat caggatctta tagaccgagg
     1921 atggagaaga agtgggaaat atgtgtacaa acctgtcatg gatcaaacat gctgtcctca
     1981 gtatacaata aggtgtcatc ctttacagtt tcagccatca aaatctcaca agaaagtttt
     2041 gaaaaaaatg ctgaaatttc tggctaaagg agagatctcg aaaggcaatt gtgaggatga
     2101 gcccatggat tctacagtgg aggatgctgt tgacggtgac tttgcattaa ttaacaagct
     2161 ggatataaag tgtgatctca aaacactcag tgacctcaaa ggaagcatag agagtgaaga
     2221 gaaggagaaa gaaaagagta taaagaaaga agggtctaaa gaattcattc atccacaatc
     2281 tatagaggag aagttgggct ctggtgaacc atcacatcca atcaaagttc atattggtcc
     2341 taagccaggc aaaggggctg acttgagtaa gcctccatgt cggaaagcaa gggaaatgag
     2401 gaaagaaagg caaagattaa aacggatgca gcaggcctca gctgcagcct cggaggctca
     2461 aggtcagcca gtctgtttgt taccaaaggc taaatccaac cagcccaagt cactggaaga
     2521 tttgattttt caatctttac cagaaaatgc atcgcacaag ttagaggtga ggctagtacc
     2581 tgcctccttt gaggacccag agttcaactc atccttcaac cagtcctttt ctttatatac
     2641 caagtatcaa gtggctatac accaggaagc acctgaaata tgtgaaaagt ctgagtttac
     2701 aagattcctt tgcagctcac cattggaggc agagcaccct gctgatggac cagaatgtgg
     2761 ttatggctcc tttcaccagc agtactggct cgatgggaag atcattgctg tgggggtgtt
     2821 agacattctc ccgtactgtg tgtcttctgt gtatctctac tacgatcctg attattcatt
     2881 tctgtctttg ggtgtctatt cagcattaag agaaattgct tttactagac aactgcatga
     2941 gaaaacatcg caactcagct attattatat gggtttctac attcattcct gtcccaagat
     3001 gagatacaag ggtcaatata gaccttctga tttgctgtgt cctgagacgt atgtctgggt
     3061 gcccattgag cagtgcctgc cttccctgga caactccaag tactgccgtt tcaaccagga
     3121 ccccgaagca gaggatgaag gacgcagtaa ggaacttgac cgactaaggg tgtttcacag
     3181 acgatctgcc atgccttacg gtgtttataa gaatcaccaa gaagacccaa gtgaggaggc
     3241 tggtgtgctg gagtatgcaa acctcgtagg acagaagtgc tcggagagga tgctgctgtt
     3301 cagacactga ggatccaagc ttagatccgg ctgctaacaa agcccgaaag gaagctgagt
     3361 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
     3421 tgaggggttt tttgctgaaa ggaggaacta tatccggata tcccgcaaga ggcccggcag
     3481 taccggcata accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat
     3541 gagcgcattg ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa
     3601 actaccgcat taaagctaat tcttgaagac gaaagggcct cgtgatacgc ctatttttat
     3661 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     3721 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3781 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3841 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3901 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3961 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4021 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     4081 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4141 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4201 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4261 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4321 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg
     4381 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4441 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4501 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4561 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4621 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4681 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     4741 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4801 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4861 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4921 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4981 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     5041 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5101 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5161 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5221 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5281 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5341 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5401 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5461 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5521 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     5581 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     5641 ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac
     5701 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     5761 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     5821 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     5881 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     5941 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     6001 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     6061 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
     6121 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
     6181 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
     6241 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
     6301 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
     6361 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
     6421 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
     6481 cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg tggccaggac
     6541 ccaacgctgc ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg
     6601 ttctgccaag ggttggtttg cgcattcaca gttctccgca agaattgatt ggctccaatt
     6661 cttggagtgg tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc gaggtggccc
     6721 ggctccatgc accgcgacgc aacgcgggga ggcagacaag gtatagggcg gcgcctacaa
     6781 tccatgccaa cccgttccat gtgctcgccg aggcggcata aatcgccgtg acgatcagcg
     6841 gtccaatgat cgaagttagg ctggtaagag ccgcgagcga tccttgaagc tgtccctgat
     6901 ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc gggcatcccg atgccgccgg
     6961 aagcgagaag aatcataatg gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga
     7021 cgtagcccag cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt
     7081 tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa
     7141 gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga
     7201 gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga
     7261 cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca
     7321 tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac
     7381 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     7441 cggggagagg cg
//
