LOCUS       sequence-217496-        4384 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli expression vector for IHFa E10A + 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: pAW203
FEATURES             Location/Qualifiers
     source          1..4384
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    54..78
                     /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        487..505
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    506..530
                     /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(873..891)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      887..934
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1155..1814)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1729..1748
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1815..1917)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2309..2326)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2443..2988)
                     /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(2523..2542)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3213..4295)
                     /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     4257..4276
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4296..4373)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cgctctccct tatgcgactc ctgcattagg aaattaatac gactcactat aggggaattg
       61 tgagcggata acaattcccc tgtagaaata attttgttta actttaataa ggagatatac
      121 catggcgctt acaaaagctg aaatgtcagc atatctgttt gataagcttg ggcttagcaa
      181 gcgggatgcc aaagaactgg ttgaactgtt tttcgaagag atccgtcgcg ctctggaaaa
      241 cggcgaacag gtgaaactct ctggttttgg taacttcgat ctgcgtgata agaatcaacg
      301 cccgggacgt aacccgaaaa cgggcgagga tattcccatt acagcacggc gcgtggtgac
      361 cttcagaccc gggcagaagt taaaaagccg ggtcgaaaac gcttcgccca aagacgagta
      421 agcggccgca taatgcttaa gtcgaacaga aagtaatcgt attgtacacg gccgcataat
      481 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc ccatcttagt
      541 atattagtta agtataagaa ggagatatac atatgaccaa gtcagaattg atagaaagac
      601 ttgccaccca gcaatcgcac attcccgcca agacggttga agatgcagta aaagagatgc
      661 tggagcatat ggcctcgact cttgcgcagg gcgagcgtat tgaaatccgc ggtttcggca
      721 gtttctcttt gcactaccgc gcaccacgta ccggacgtaa tccgaagact ggcgataaag
      781 tagaactgga aggaaaatac gttcctcact ttaaacctgg taaagaactg cgcgatcgcg
      841 ccaatattta cggttaacct aggctgctgc caccgctgag caataactag cataacccct
      901 tggggcctct aaacgggtct tgaggggttt tttgctgaaa cctcaggcat ttgagaagca
      961 cacggtcaca ctgcttccgg tagtcaataa accggtaaac cagcaataga cataagcggc
     1021 tatttaacga ccctgccctg aaccgacgac cgggtcgaat ttgctttcga atttctgcca
     1081 ttcatccgct tattatcact tattcaggcg tagcaaccag gcgtttaagg gcaccaataa
     1141 ctgccttaaa aaaattacgc cccgccctgc cactcatcgc agtactgttg taattcatta
     1201 agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc
     1261 atcagcacct tgtcgccttg cgtataatat ttgcccatag tgaaaacggg ggcgaagaag
     1321 ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag
     1381 acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac
     1441 gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag
     1501 agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc
     1561 catatcacca gctcaccgtc tttcattgcc atacggaact ccggatgagc attcatcagg
     1621 cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt
     1681 aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga
     1741 aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg
     1801 atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg
     1861 cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaacg
     1921 tctcattttc gccaaaagtt ggcccagggc ttcccggtat caacagggac accaggattt
     1981 atttattctg cgaagtgatc ttccgtcaca ggtatttatt cggcgcaaag tgcgtcgggt
     2041 gatgctgcca acttactgat ttagtgtatg atggtgtttt tgaggtgctc cagtggcttc
     2101 tgtttctatc agctgtccct cctgttcagc tactgacggg gtggtgcgta acggcaaaag
     2161 caccgccgga catcagcgct agcggagtgt atactggctt actatgttgg cactgatgag
     2221 ggtgtcagtg aagtgcttca tgtggcagga gaaaaaaggc tgcaccggtg cgtcagcaga
     2281 atatgtgata caggatatat tccgcttcct cgctcactga ctcgctacgc tcggtcgttc
     2341 gactgcggcg agcggaaatg gcttacgaac ggggcggaga tttcctggaa gatgccagga
     2401 agatacttaa cagggaagtg agagggccgc ggcaaagccg tttttccata ggctccgccc
     2461 ccctgacaag catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc cgacaggact
     2521 ataaagatac caggcgtttc cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt
     2581 tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg tctcattcca cgcctgacac
     2641 tcagttccgg gtaggcagtt cgctccaagc tggactgtat gcacgaaccc cccgttcagt
     2701 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggaa agacatgcaa
     2761 aagcaccact ggcagcagcc actggtaatt gatttagagg agttagtctt gaagtcatgc
     2821 gccggttaag gctaaactga aaggacaagt tttggtgact gcgctcctcc aagccagtta
     2881 cctcggttca aagagttggt agctcagaga accttcgaaa aaccgccctg caaggcggtt
     2941 ttttcgtttt cagagcaaga gattacgcgc agaccaaaac gatctcaaga agatcatctt
     3001 attaatcaga taaaatattt ctagatttca gtgcaattta tctcttcaaa tgtagcacct
     3061 gaagtcagcc ccatacgata taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc
     3121 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat
     3181 gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3241 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3301 ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc
     3361 gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa
     3421 tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat
     3481 cccactaccg agatgtccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg
     3541 cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc
     3601 atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc
     3661 ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag
     3721 acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc
     3781 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg
     3841 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca
     3901 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg
     3961 tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg
     4021 gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg
     4081 gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc
     4141 acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc
     4201 gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca
     4261 tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct
     4321 tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc
     4381 tcga
//
