LOCUS       sequence-218810-        4384 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli expression vector for IHFa F13A + 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: pAW204
FEATURES             Location/Qualifiers
     source          1..4384
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..43
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    44..68
                     /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        477..495
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    496..520
                     /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(863..881)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      877..924
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1145..1804)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1719..1738
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1805..1907)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2299..2316)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2433..2978)
                     /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(2513..2532)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3203..4285)
                     /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     4247..4266
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4286..4363)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 tatgcgactc ctgcattagg aaattaatac gactcactat aggggaattg tgagcggata
       61 acaattcccc tgtagaaata attttgttta actttaataa ggagatatac catggcgctt
      121 acaaaagctg aaatgtcaga atatctggca gataagcttg ggcttagcaa gcgggatgcc
      181 aaagaactgg ttgaactgtt tttcgaagag atccgtcgcg ctctggaaaa cggcgaacag
      241 gtgaaactct ctggttttgg taacttcgat ctgcgtgata agaatcaacg cccgggacgt
      301 aacccgaaaa cgggcgagga tattcccatt acagcacggc gcgtggtgac cttcagaccc
      361 gggcagaagt taaaaagccg ggtcgaaaac gcttcgccca aagacgagta agcggccgca
      421 taatgcttaa gtcgaacaga aagtaatcgt attgtacacg gccgcataat cgaaattaat
      481 acgactcact ataggggaat tgtgagcgga taacaattcc ccatcttagt atattagtta
      541 agtataagaa ggagatatac atatgaccaa gtcagaattg atagaaagac ttgccaccca
      601 gcaatcgcac attcccgcca agacggttga agatgcagta aaagagatgc tggagcatat
      661 ggcctcgact cttgcgcagg gcgagcgtat tgaaatccgc ggtttcggca gtttctcttt
      721 gcactaccgc gcaccacgta ccggacgtaa tccgaagact ggcgataaag tagaactgga
      781 aggaaaatac gttcctcact ttaaacctgg taaagaactg cgcgatcgcg ccaatattta
      841 cggttaacct aggctgctgc caccgctgag caataactag cataacccct tggggcctct
      901 aaacgggtct tgaggggttt tttgctgaaa cctcaggcat ttgagaagca cacggtcaca
      961 ctgcttccgg tagtcaataa accggtaaac cagcaataga cataagcggc tatttaacga
     1021 ccctgccctg aaccgacgac cgggtcgaat ttgctttcga atttctgcca ttcatccgct
     1081 tattatcact tattcaggcg tagcaaccag gcgtttaagg gcaccaataa ctgccttaaa
     1141 aaaattacgc cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc
     1201 cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct
     1261 tgtcgccttg cgtataatat ttgcccatag tgaaaacggg ggcgaagaag ttgtccatat
     1321 tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca
     1381 tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt
     1441 gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa
     1501 acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca
     1561 gctcaccgtc tttcattgcc atacggaact ccggatgagc attcatcagg cgggcaagaa
     1621 tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg
     1681 taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa
     1741 aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg atttttttct
     1801 ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     1861 atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaacg tctcattttc
     1921 gccaaaagtt ggcccagggc ttcccggtat caacagggac accaggattt atttattctg
     1981 cgaagtgatc ttccgtcaca ggtatttatt cggcgcaaag tgcgtcgggt gatgctgcca
     2041 acttactgat ttagtgtatg atggtgtttt tgaggtgctc cagtggcttc tgtttctatc
     2101 agctgtccct cctgttcagc tactgacggg gtggtgcgta acggcaaaag caccgccgga
     2161 catcagcgct agcggagtgt atactggctt actatgttgg cactgatgag ggtgtcagtg
     2221 aagtgcttca tgtggcagga gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata
     2281 caggatatat tccgcttcct cgctcactga ctcgctacgc tcggtcgttc gactgcggcg
     2341 agcggaaatg gcttacgaac ggggcggaga tttcctggaa gatgccagga agatacttaa
     2401 cagggaagtg agagggccgc ggcaaagccg tttttccata ggctccgccc ccctgacaag
     2461 catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc cgacaggact ataaagatac
     2521 caggcgtttc cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc
     2581 ggtgtcattc cgctgttatg gccgcgtttg tctcattcca cgcctgacac tcagttccgg
     2641 gtaggcagtt cgctccaagc tggactgtat gcacgaaccc cccgttcagt ccgaccgctg
     2701 cgccttatcc ggtaactatc gtcttgagtc caacccggaa agacatgcaa aagcaccact
     2761 ggcagcagcc actggtaatt gatttagagg agttagtctt gaagtcatgc gccggttaag
     2821 gctaaactga aaggacaagt tttggtgact gcgctcctcc aagccagtta cctcggttca
     2881 aagagttggt agctcagaga accttcgaaa aaccgccctg caaggcggtt ttttcgtttt
     2941 cagagcaaga gattacgcgc agaccaaaac gatctcaaga agatcatctt attaatcaga
     3001 taaaatattt ctagatttca gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc
     3061 ccatacgata taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc ggaaggagct
     3121 gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta
     3181 acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3241 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg
     3301 tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct
     3361 gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga
     3421 tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg
     3481 agatgtccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca
     3541 tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg
     3601 tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt
     3661 gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta
     3721 atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca
     3781 gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat
     3841 caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat
     3901 ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg
     3961 ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt
     4021 gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg
     4081 aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg
     4141 gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt
     4201 ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga
     4261 catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct
     4321 atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct
     4381 ccct
//
