LOCUS       sequence-312933-        6058 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Amp resistant IPTG inducible 6xHis-TEV-ABLE protein for E. coli 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6058)
  AUTHORS   Polizzi NF, DeGrado WF
  TITLE     A defined structural unit enables de novo design of 
            small-molecule-binding proteins.
  JOURNAL   Science. 2020 Sep 4;369(6508):1227-1233. doi: 
            10.1126/science.abb8330.
  PUBMED    32883865
REFERENCE   2  (bases 1 to 6058)
  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"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2020-09-4- 4"; volume: "369"; issue: 
            "6508"; pages: "1227-1233"
COMMENT     Sequence Label: pet11a-ABLE
FEATURES             Location/Qualifiers
     source          1..6058
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             92..109
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             110..130
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(559..577)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      573..620
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(795..800,801..817,818..823))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(850..868)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        936..1040
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1041..1109,1110..1901)
                     /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(1259..1278)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2072..2660
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2561..2580
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2814..2831
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2846..2985
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2911..2930)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3071..3093
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3087..3278)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4590..5672)
                     /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"
     primer_bind     5634..5653
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5673..5750)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5956..5975
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gcaccaccac caccaccacg aaaacctgta
      121 cttccagagc gtgaagagcg agtatgcgga agctgcggcg gttggtcaag aagcggtggc
      181 ggttttcaac accatgaagg cggcgtttca gaacggcgat aaagaggcgg ttgcgcaata
      241 cctggcgcgt ctggcgagcc tgtatacccg tcacgaggaa ctgctgaacc gtatcctgga
      301 aaaggcgcgt cgtgagggta acaaagaagc ggtgaccctg atgaacgagt tcaccgcgac
      361 ctttcagacc ggcaagagca ttttcaacgc gatggttgcg gcgtttaaaa acggcgacga
      421 tgacagcttt gagagctacc tgcaggcgct ggaaaaggtg accgcgaaag gcgagaccct
      481 ggcggaccaa atcgcgaaag cgctgtaagg atccggctgc taacaaagcc cgaaaggaag
      541 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
      601 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatatccc gcaagaggcc
      661 cggcagtacc ggcataacca agcctatgcc tacagcatcc agggtgacgg tgccgaggat
      721 gacgatgagc gcattgttag atttcataca cggtgcctga ctgcgttagc aatttaactg
      781 tgataaacta ccgcattaaa gcttatcgat gataagctgt caaacatgag aattcttgaa
      841 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
      901 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      961 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1021 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     1081 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     1141 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     1201 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     1261 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     1321 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     1381 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     1441 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     1501 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     1561 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
     1621 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     1681 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     1741 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     1801 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1861 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1921 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1981 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     2041 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2101 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2161 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     2221 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2281 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2341 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2401 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2461 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2521 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     2581 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     2641 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     2701 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     2761 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     2821 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     2881 gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     2941 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
     3001 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     3061 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     3121 aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt
     3181 tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag
     3241 cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga
     3301 tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt
     3361 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg
     3421 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat
     3481 gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg
     3541 cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat
     3601 gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc
     3661 ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac
     3721 aggagcacga tcatgcgcac ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg
     3781 cggctgctgg agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc
     3841 acagttctcc gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg
     3901 tgccgccggc ttccattcag gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg
     3961 ggaggcagac aaggtatagg gcggcgccta caatccatgc caacccgttc catgtgctcg
     4021 ccgaggcggc ataaatcgcc gtgacgatca gcggtccaat gatcgaagtt aggctggtaa
     4081 gagccgcgag cgatccttga agctgtccct gatggtcgtc atctacctgc ctggacagca
     4141 tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atggggaagg
     4201 ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc
     4261 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt
     4321 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc
     4381 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt
     4441 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga
     4501 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
     4561 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4621 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4681 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
     4741 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
     4801 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
     4861 actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
     4921 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
     4981 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     5041 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     5101 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     5161 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     5221 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     5281 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     5341 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     5401 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
     5461 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
     5521 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
     5581 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
     5641 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     5701 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg
     5761 acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt
     5821 gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc
     5881 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     5941 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     6001 ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc cgcgaaat
//
