LOCUS       sequence-220897-        4384 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli expression vector for IHFa K20A + IHFb.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4384)
  AUTHORS   Wright AV, Liu JJ, Knott GJ, Doxzen KW, Nogales E, Doudna JA
  TITLE     Structures of the CRISPR genome integration complex.
  JOURNAL   Science. 2017 Sep 15;357(6356):1113-1118. doi: 
            10.1126/science.aao0679. Epub 2017 Jul 20.
  PUBMED    28729350
REFERENCE   2  (bases 1 to 4384)
  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.aao0679"; journalName: "Science"; date: 
            "2017-09-15- 15"; volume: "357"; issue: "6356"; pages: "1113-1118"
COMMENT     Sequence Label: pAW207
FEATURES             Location/Qualifiers
     source          1..4384
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        41..59
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    60..84
                     /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)."
     promoter        493..511
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    512..536
                     /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)."
     primer_bind     complement(879..897)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      893..940
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1161..1820)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1735..1754
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1821..1923)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2315..2332)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2449..2994)
                     /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(2529..2548)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3219..4301)
                     /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     4263..4282
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4302..4379)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 tctcgacgct ctcccttatg cgactcctgc attaggaaat taatacgact cactataggg
       61 gaattgtgag cggataacaa ttcccctgta gaaataattt tgtttaactt taataaggag
      121 atataccatg gcgcttacaa aagctgaaat gtcagaatat ctgtttgata agcttgggct
      181 tagcgcacgg gatgccaaag aactggttga actgtttttc gaagagatcc gtcgcgctct
      241 ggaaaacggc gaacaggtga aactctctgg ttttggtaac ttcgatctgc gtgataagaa
      301 tcaacgcccg ggacgtaacc cgaaaacggg cgaggatatt cccattacag cacggcgcgt
      361 ggtgaccttc agacccgggc agaagttaaa aagccgggtc gaaaacgctt cgcccaaaga
      421 cgagtaagcg gccgcataat gcttaagtcg aacagaaagt aatcgtattg tacacggccg
      481 cataatcgaa attaatacga ctcactatag gggaattgtg agcggataac aattccccat
      541 cttagtatat tagttaagta taagaaggag atatacatat gaccaagtca gaattgatag
      601 aaagacttgc cacccagcaa tcgcacattc ccgccaagac ggttgaagat gcagtaaaag
      661 agatgctgga gcatatggcc tcgactcttg cgcagggcga gcgtattgaa atccgcggtt
      721 tcggcagttt ctctttgcac taccgcgcac cacgtaccgg acgtaatccg aagactggcg
      781 ataaagtaga actggaagga aaatacgttc ctcactttaa acctggtaaa gaactgcgcg
      841 atcgcgccaa tatttacggt taacctaggc tgctgccacc gctgagcaat aactagcata
      901 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaacctc aggcatttga
      961 gaagcacacg gtcacactgc ttccggtagt caataaaccg gtaaaccagc aatagacata
     1021 agcggctatt taacgaccct gccctgaacc gacgaccggg tcgaatttgc tttcgaattt
     1081 ctgccattca tccgcttatt atcacttatt caggcgtagc aaccaggcgt ttaagggcac
     1141 caataactgc cttaaaaaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat
     1201 tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc
     1261 agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatagtgaa aacgggggcg
     1321 aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg
     1381 gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg
     1441 taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca
     1501 ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca
     1561 ctatcccata tcaccagctc accgtctttc attgccatac ggaactccgg atgagcattc
     1621 atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg
     1681 gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact
     1741 gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat
     1801 ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa
     1861 aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga
     1921 tcaacgtctc attttcgcca aaagttggcc cagggcttcc cggtatcaac agggacacca
     1981 ggatttattt attctgcgaa gtgatcttcc gtcacaggta tttattcggc gcaaagtgcg
     2041 tcgggtgatg ctgccaactt actgatttag tgtatgatgg tgtttttgag gtgctccagt
     2101 ggcttctgtt tctatcagct gtccctcctg ttcagctact gacggggtgg tgcgtaacgg
     2161 caaaagcacc gccggacatc agcgctagcg gagtgtatac tggcttacta tgttggcact
     2221 gatgagggtg tcagtgaagt gcttcatgtg gcaggagaaa aaaggctgca ccggtgcgtc
     2281 agcagaatat gtgatacagg atatattccg cttcctcgct cactgactcg ctacgctcgg
     2341 tcgttcgact gcggcgagcg gaaatggctt acgaacgggg cggagatttc ctggaagatg
     2401 ccaggaagat acttaacagg gaagtgagag ggccgcggca aagccgtttt tccataggct
     2461 ccgcccccct gacaagcatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac
     2521 aggactataa agataccagg cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc
     2581 tgcctttcgg tttaccggtg tcattccgct gttatggccg cgtttgtctc attccacgcc
     2641 tgacactcag ttccgggtag gcagttcgct ccaagctgga ctgtatgcac gaaccccccg
     2701 ttcagtccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggaaagac
     2761 atgcaaaagc accactggca gcagccactg gtaattgatt tagaggagtt agtcttgaag
     2821 tcatgcgccg gttaaggcta aactgaaagg acaagttttg gtgactgcgc tcctccaagc
     2881 cagttacctc ggttcaaaga gttggtagct cagagaacct tcgaaaaacc gccctgcaag
     2941 gcggtttttt cgttttcaga gcaagagatt acgcgcagac caaaacgatc tcaagaagat
     3001 catcttatta atcagataaa atatttctag atttcagtgc aatttatctc ttcaaatgta
     3061 gcacctgaag tcagccccat acgatataag ttgtaattct catgttagtc atgccccgcg
     3121 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg
     3181 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
     3241 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     3301 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
     3361 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
     3421 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
     3481 tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt aatggcgcgc
     3541 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
     3601 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
     3661 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
     3721 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
     3781 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
     3841 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
     3901 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
     3961 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
     4021 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
     4081 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
     4141 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
     4201 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
     4261 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
     4321 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc
     4381 ggga
//
