LOCUS       Exported                5679 bp ds-DNA     circular UNK 30-JAN-2025
DEFINITION  Validation set of 96 E. coli-Expressed, Single-Domain Proteins with 
            Defined Structural and Biochemical Features.
ACCESSION   pAIP4
VERSION     .
KEYWORDS    .
SOURCE      .
  ORGANISM  .
REFERENCE   1  (bases 1 to 5679)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jan 30, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5679
                     /organism="."
                     /mol_type="genomic DNA"
     CDS             19..36
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     terminator      103..150
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff"
     rep_origin      187..642
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     CDS             complement(735..1550)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /note="color: #ccffcc"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      1672..2260
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     misc_feature    2446..2588
                     /label=bom
                     /note="basis of mobility region from pBR322"
                     /note="color: #a6acb3"
     CDS             complement(2690..2881)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /note="color: #993366"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3690..4772)
                     /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)."
                     /note="color: #993366"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4773..4850)
                     /gene="lacI"
                     /label=lacI promoter
                     /note="color: #ffffff; direction: LEFT"
     promoter        5163..5181
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     protein_bind    5182..5206
                     /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)."
                     /note="color: #31849b"
     RBS             5221..5243
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
                     /note="color: #a6acb3"
     CDS             5263..5292
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHHHHHH"
ORIGIN
        1 ggttcgtaat gactcgagca ccaccaccac caccactgag atccggctgc taacaaagcc
       61 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
      121 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg
      181 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
      241 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
      301 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
      361 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
      421 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
      481 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
      541 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
      601 aatttaacgc gaattttaac aaaatattaa cgtttacaat ttcaggtggc acttttcggg
      661 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
      721 tcatgaatta attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata
      781 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
      841 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
      901 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
      961 ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact
     1021 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta
     1081 ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta
     1141 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
     1201 cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg
     1261 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
     1321 tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg
     1381 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca
     1441 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
     1501 gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa tatggctcat aacacccctt
     1561 gtattactgt ttatgtaagc agacagtttt attgttcatg accaaaatcc cttaacgtga
     1621 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     1681 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     1741 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     1801 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     1861 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     1921 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     1981 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     2041 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     2101 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     2161 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     2221 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     2281 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     2341 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     2401 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     2461 tctccttacg catctgtgcg gtatttcaca ccgcatatat ggtgcactct cagtacaatc
     2521 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     2581 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     2641 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     2701 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     2761 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     2821 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     2881 ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag
     2941 agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg
     3001 agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat
     3061 gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga
     3121 tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa
     3181 cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt
     3241 cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc
     3301 agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggg gccgccatgc
     3361 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt
     3421 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc
     3481 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt
     3541 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga
     3601 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
     3661 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3721 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3781 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
     3841 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
     3901 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
     3961 actaccgaga tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
     4021 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
     4081 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     4141 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     4201 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     4261 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     4321 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     4381 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     4441 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     4501 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
     4561 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
     4621 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
     4681 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
     4741 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     4801 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg
     4861 acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt
     4921 gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc
     4981 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     5041 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     5101 ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atcgatctcg atcccgcgaa
     5161 attaatacga ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat
     5221 tttgtttaac tttaagaagg agatatacat atggcgaaga ttcatcacca tcatcaccat
     5281 caccatcacc acggtagcct gcaagattcg gaagtcaacc aagaagcaaa gccggaagtc
     5341 aagccggaag tgaaaccgga aacccatatt aacctgaaag ttagtgacgg cagctctgaa
     5401 attttcttta agatcaaaaa gaccacgccg ctgcgtcgcc tgatggaagc gtttgccaaa
     5461 cgtcagggca aggaaatgga tagcctgcgt ttcctgtatg acggtattcg catccaggca
     5521 gatcaagcgc cggaagacct ggacatggaa gacaacgaca tcattgaagc ccaccgtgaa
     5581 cagattggcg gctctggaga gaccggagcg agacgggagt cgtcttcgct ttccagatct
     5641 gataacttgt gaagacgacc atcgtctcac agtggtctc
//
