LOCUS       sequence-220894-        4384 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli expression vector for IHFa K15A + 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: pAW206
FEATURES             Location/Qualifiers
     source          1..4384
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     343..362
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(382..459)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        505..523
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    524..548
                     /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        957..975
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    976..1000
                     /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(1343..1361)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1357..1404
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1625..2284)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2199..2218
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2285..2387)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2779..2796)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2913..3458)
                     /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(2993..3012)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3683..4384,1..381))
                     /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 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
       61 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
      121 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
      181 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
      241 gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
      301 ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc
      361 gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca
      421 tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct
      481 tatgcgactc ctgcattagg aaattaatac gactcactat aggggaattg tgagcggata
      541 acaattcccc tgtagaaata attttgttta actttaataa ggagatatac catggcgctt
      601 acaaaagctg aaatgtcaga atatctgttt gatgcacttg ggcttagcaa gcgggatgcc
      661 aaagaactgg ttgaactgtt tttcgaagag atccgtcgcg ctctggaaaa cggcgaacag
      721 gtgaaactct ctggttttgg taacttcgat ctgcgtgata agaatcaacg cccgggacgt
      781 aacccgaaaa cgggcgagga tattcccatt acagcacggc gcgtggtgac cttcagaccc
      841 gggcagaagt taaaaagccg ggtcgaaaac gcttcgccca aagacgagta agcggccgca
      901 taatgcttaa gtcgaacaga aagtaatcgt attgtacacg gccgcataat cgaaattaat
      961 acgactcact ataggggaat tgtgagcgga taacaattcc ccatcttagt atattagtta
     1021 agtataagaa ggagatatac atatgaccaa gtcagaattg atagaaagac ttgccaccca
     1081 gcaatcgcac attcccgcca agacggttga agatgcagta aaagagatgc tggagcatat
     1141 ggcctcgact cttgcgcagg gcgagcgtat tgaaatccgc ggtttcggca gtttctcttt
     1201 gcactaccgc gcaccacgta ccggacgtaa tccgaagact ggcgataaag tagaactgga
     1261 aggaaaatac gttcctcact ttaaacctgg taaagaactg cgcgatcgcg ccaatattta
     1321 cggttaacct aggctgctgc caccgctgag caataactag cataacccct tggggcctct
     1381 aaacgggtct tgaggggttt tttgctgaaa cctcaggcat ttgagaagca cacggtcaca
     1441 ctgcttccgg tagtcaataa accggtaaac cagcaataga cataagcggc tatttaacga
     1501 ccctgccctg aaccgacgac cgggtcgaat ttgctttcga atttctgcca ttcatccgct
     1561 tattatcact tattcaggcg tagcaaccag gcgtttaagg gcaccaataa ctgccttaaa
     1621 aaaattacgc cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc
     1681 cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct
     1741 tgtcgccttg cgtataatat ttgcccatag tgaaaacggg ggcgaagaag ttgtccatat
     1801 tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca
     1861 tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt
     1921 gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa
     1981 acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca
     2041 gctcaccgtc tttcattgcc atacggaact ccggatgagc attcatcagg cgggcaagaa
     2101 tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg
     2161 taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa
     2221 aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg atttttttct
     2281 ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     2341 atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaacg tctcattttc
     2401 gccaaaagtt ggcccagggc ttcccggtat caacagggac accaggattt atttattctg
     2461 cgaagtgatc ttccgtcaca ggtatttatt cggcgcaaag tgcgtcgggt gatgctgcca
     2521 acttactgat ttagtgtatg atggtgtttt tgaggtgctc cagtggcttc tgtttctatc
     2581 agctgtccct cctgttcagc tactgacggg gtggtgcgta acggcaaaag caccgccgga
     2641 catcagcgct agcggagtgt atactggctt actatgttgg cactgatgag ggtgtcagtg
     2701 aagtgcttca tgtggcagga gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata
     2761 caggatatat tccgcttcct cgctcactga ctcgctacgc tcggtcgttc gactgcggcg
     2821 agcggaaatg gcttacgaac ggggcggaga tttcctggaa gatgccagga agatacttaa
     2881 cagggaagtg agagggccgc ggcaaagccg tttttccata ggctccgccc ccctgacaag
     2941 catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc cgacaggact ataaagatac
     3001 caggcgtttc cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc
     3061 ggtgtcattc cgctgttatg gccgcgtttg tctcattcca cgcctgacac tcagttccgg
     3121 gtaggcagtt cgctccaagc tggactgtat gcacgaaccc cccgttcagt ccgaccgctg
     3181 cgccttatcc ggtaactatc gtcttgagtc caacccggaa agacatgcaa aagcaccact
     3241 ggcagcagcc actggtaatt gatttagagg agttagtctt gaagtcatgc gccggttaag
     3301 gctaaactga aaggacaagt tttggtgact gcgctcctcc aagccagtta cctcggttca
     3361 aagagttggt agctcagaga accttcgaaa aaccgccctg caaggcggtt ttttcgtttt
     3421 cagagcaaga gattacgcgc agaccaaaac gatctcaaga agatcatctt attaatcaga
     3481 taaaatattt ctagatttca gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc
     3541 ccatacgata taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc ggaaggagct
     3601 gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta
     3661 acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3721 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg
     3781 tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct
     3841 gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga
     3901 tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg
     3961 agatgtccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca
     4021 tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg
     4081 tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt
     4141 gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta
     4201 atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca
     4261 gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat
     4321 caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat
     4381 ccag
//
