LOCUS       sequence-220892-        4384 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli expression vector for IHFa D14A + 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: pAW205
FEATURES             Location/Qualifiers
     source          1..4384
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        38..56
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    57..81
                     /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        490..508
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    509..533
                     /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(876..894)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      890..937
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1158..1817)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1732..1751
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1818..1920)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2312..2329)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2446..2991)
                     /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(2526..2545)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3216..4298)
                     /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     4260..4279
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4299..4376)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cgacgctctc ccttatgcga ctcctgcatt aggaaattaa tacgactcac tataggggaa
       61 ttgtgagcgg ataacaattc ccctgtagaa ataattttgt ttaactttaa taaggagata
      121 taccatggcg cttacaaaag ctgaaatgtc agaatatctg tttgcaaagc ttgggcttag
      181 caagcgggat gccaaagaac tggttgaact gtttttcgaa gagatccgtc gcgctctgga
      241 aaacggcgaa caggtgaaac tctctggttt tggtaacttc gatctgcgtg ataagaatca
      301 acgcccggga cgtaacccga aaacgggcga ggatattccc attacagcac ggcgcgtggt
      361 gaccttcaga cccgggcaga agttaaaaag ccgggtcgaa aacgcttcgc ccaaagacga
      421 gtaagcggcc gcataatgct taagtcgaac agaaagtaat cgtattgtac acggccgcat
      481 aatcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tccccatctt
      541 agtatattag ttaagtataa gaaggagata tacatatgac caagtcagaa ttgatagaaa
      601 gacttgccac ccagcaatcg cacattcccg ccaagacggt tgaagatgca gtaaaagaga
      661 tgctggagca tatggcctcg actcttgcgc agggcgagcg tattgaaatc cgcggtttcg
      721 gcagtttctc tttgcactac cgcgcaccac gtaccggacg taatccgaag actggcgata
      781 aagtagaact ggaaggaaaa tacgttcctc actttaaacc tggtaaagaa ctgcgcgatc
      841 gcgccaatat ttacggttaa cctaggctgc tgccaccgct gagcaataac tagcataacc
      901 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaacctcagg catttgagaa
      961 gcacacggtc acactgcttc cggtagtcaa taaaccggta aaccagcaat agacataagc
     1021 ggctatttaa cgaccctgcc ctgaaccgac gaccgggtcg aatttgcttt cgaatttctg
     1081 ccattcatcc gcttattatc acttattcag gcgtagcaac caggcgttta agggcaccaa
     1141 taactgcctt aaaaaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca
     1201 ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc
     1261 ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tagtgaaaac gggggcgaag
     1321 aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca gggattggct
     1381 gagacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa
     1441 cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc
     1501 cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta
     1561 tcccatatca ccagctcacc gtctttcatt gccatacgga actccggatg agcattcatc
     1621 aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc
     1681 tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac
     1741 tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca
     1801 gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat
     1861 acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca
     1921 acgtctcatt ttcgccaaaa gttggcccag ggcttcccgg tatcaacagg gacaccagga
     1981 tttatttatt ctgcgaagtg atcttccgtc acaggtattt attcggcgca aagtgcgtcg
     2041 ggtgatgctg ccaacttact gatttagtgt atgatggtgt ttttgaggtg ctccagtggc
     2101 ttctgtttct atcagctgtc cctcctgttc agctactgac ggggtggtgc gtaacggcaa
     2161 aagcaccgcc ggacatcagc gctagcggag tgtatactgg cttactatgt tggcactgat
     2221 gagggtgtca gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc
     2281 agaatatgtg atacaggata tattccgctt cctcgctcac tgactcgcta cgctcggtcg
     2341 ttcgactgcg gcgagcggaa atggcttacg aacggggcgg agatttcctg gaagatgcca
     2401 ggaagatact taacagggaa gtgagagggc cgcggcaaag ccgtttttcc ataggctccg
     2461 cccccctgac aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa acccgacagg
     2521 actataaaga taccaggcgt ttccccctgg cggctccctc gtgcgctctc ctgttcctgc
     2581 ctttcggttt accggtgtca ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga
     2641 cactcagttc cgggtaggca gttcgctcca agctggactg tatgcacgaa ccccccgttc
     2701 agtccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gaaagacatg
     2761 caaaagcacc actggcagca gccactggta attgatttag aggagttagt cttgaagtca
     2821 tgcgccggtt aaggctaaac tgaaaggaca agttttggtg actgcgctcc tccaagccag
     2881 ttacctcggt tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg
     2941 gttttttcgt tttcagagca agagattacg cgcagaccaa aacgatctca agaagatcat
     3001 cttattaatc agataaaata tttctagatt tcagtgcaat ttatctcttc aaatgtagca
     3061 cctgaagtca gccccatacg atataagttg taattctcat gttagtcatg ccccgcgccc
     3121 accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct
     3181 aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3241 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3301 ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc
     3361 accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga
     3421 aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg
     3481 tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt
     3541 gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc
     3601 agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct
     3661 atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc
     3721 gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga
     3781 tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc
     3841 tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg
     3901 gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga
     3961 ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg
     4021 ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc
     4081 agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt
     4141 gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt
     4201 ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg
     4261 gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc
     4321 tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg
     4381 atct
//
