LOCUS       sequence-213826-        4382 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 4382)
  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 4382)
  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..4382
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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        436..454
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    455..479
                     /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(822..840)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      836..883
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1103..1762)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1677..1696
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1763..1865)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2257..2274)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2391..2935)
                     /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(2471..2490)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3160..4242)
                     /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     4204..4223
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4243..4320)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(4366..4382,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atggcgctta caaaagctga aatgtcagca tatctgtttg ataagcttgg
      121 gcttagcaag cgggatgcca aagaactggt tgaactgttt ttcgaagaga tccgtcgcgc
      181 tctggaaaac ggcgaacagg tgaaactctc tggttttggt aacttcgatc tgcgtgataa
      241 gaatcaacgc ccgggacgta acccgaaaac gggcgaggat attcccatta cagcacggcg
      301 cgtggtgacc ttcagacccg ggcagaagtt aaaaagccgg gtcgaaaacg cttcgcccaa
      361 agacgagtaa gcggccgcat aatgcttaag tcgaacagaa agtaatcgta ttgtacacgg
      421 ccgcataatc gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc
      481 catcttagta tattagttaa gtataagaag gagatataca tatgaccaag tcagaattga
      541 tagaaagact tgccacccag caatcgcaca ttcccgccaa gacggttgaa gatgcagtaa
      601 aagagatgct ggagcatatg gcctcgactc ttgcgcaggg cgagcgtatt gaaatccgcg
      661 gtttcggcag tttctctttg cactaccgcg caccacgtac cggacgtaat ccgaagactg
      721 gcgataaagt agaactggaa ggaaaatacg ttcctcactt taaacctggt aaagaactgc
      781 gcgatcgcgc caatatttac ggttaaccta ggctgctgcc accgctgagc aataactagc
      841 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaac ctcaggcatt
      901 tgagaagcac acggtcacac tgcttccggt agtcaataaa ccggtaaacc agcaatagac
      961 ataagcggct atttaacgac cctgccctga accgacgacc gggtcgaatt tgctttcgaa
     1021 tttctgccat tcatccgctt attatcactt attcaggcgt agcaccaggc gtttaagggc
     1081 accaataact gccttaaaaa aattacgccc cgccctgcca ctcatcgcag tactgttgta
     1141 attcattaag cattctgccg acatggaagc catcacagac ggcatgatga acctgaatcg
     1201 ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatagtg aaaacggggg
     1261 cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat
     1321 tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac
     1381 cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt
     1441 cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa
     1501 cactatccca tatcaccagc tcaccgtctt tcattgccat acggaactcc ggatgagcat
     1561 tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta
     1621 cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa
     1681 ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat
     1741 atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa
     1801 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     1861 gatcaacgtc tcattttcgc caaaagttgg cccagggctt cccggtatca acagggacac
     1921 caggatttat ttattctgcg aagtgatctt ccgtcacagg tatttattcg gcgcaaagtg
     1981 cgtcgggtga tgctgccaac ttactgattt agtgtatgat ggtgtttttg aggtgctcca
     2041 gtggcttctg tttctatcag ctgtccctcc tgttcagcta ctgacggggt ggtgcgtaac
     2101 ggcaaaagca ccgccggaca tcagcgctag cggagtgtat actggcttac tatgttggca
     2161 ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg caccggtgcg
     2221 tcagcagaat atgtgataca ggatatattc cgcttcctcg ctcactgact cgctacgctc
     2281 ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt tcctggaaga
     2341 tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt tttccatagg
     2401 ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg
     2461 acaggactat aaagatacca ggcgtttccc ctggcggctc cctcgtgcgc tctcctgttc
     2521 ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct cattccacgc
     2581 ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca cgaacccccc
     2641 gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggaaaga
     2701 catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt tagtcttgaa
     2761 gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg ctcctccaag
     2821 ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa
     2881 ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga
     2941 tcatcttatt aatcagataa aatatttcta gatttcagtg caatttatct cttcaaatgt
     3001 agcacctgaa gtcagcccca tacgatataa gttgtaattc tcatgttagt catgccccgc
     3061 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt
     3121 gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3181 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3241 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
     3301 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
     3361 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc
     3421 gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg
     3481 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
     3541 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
     3601 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
     3661 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
     3721 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
     3781 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
     3841 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
     3901 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     3961 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     4021 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     4081 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     4141 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     4201 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
     4261 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc
     4321 cgggatctcg acgctctccc ttatgcgact cctgcattag gaaattaata cgactcacta
     4381 ta
//
