LOCUS       sequence-228217-        7765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7765)
  AUTHORS   Deplancke B, Mukhopadhyay A, Ao W, Elewa AM, Grove CA, Martinez NJ, 
            Sequerra R, Doucette-Stamm L, Reece-Hoyes JS, Hope IA, Tissenbaum 
            HA, Mango SE, Walhout AJ
  TITLE     A gene-centered C. elegans protein-DNA interaction network.
  JOURNAL   Cell. 2006 Jun 16. 125(6):1193-205.
  PUBMED    16777607
REFERENCE   2  (bases 1 to 7765)
  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"; journalName: "Cell. 2006 
            Jun 16. 125(6):1193-205."
COMMENT     Sequence Label: pMW#2
FEATURES             Location/Qualifiers
     source          1..7765
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        25..43
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     25..42
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    113..236
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(626..931)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYHLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(728..747)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1251..1934)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     1849..1868
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1935..2037)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    2194..2293
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2445..3107
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     protein_bind    complement(3743..3767)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3817..3836)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3818..3836)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3855..3871)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3855..3871)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3868..3890)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3879..3895
                     /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        complement(join(3903..3909,3910..3927,3928..3933))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3948..3969
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(3982..5064)
                     /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     5026..5045
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5065..5142)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(5249..5266)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5420..6008)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5500..5519)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6179..6970,6971..7039))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6802..6821
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7040..7144)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7170..7624)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7307..7328)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7518..7537
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(7751..7765,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(7765,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttgaatttag gtgacactat agaagagcta tgacgtcgca
       61 tgcacgcgta cgtaagcttg gatcctctag agcggccgcc gactagtgat atcaactttg
      121 tatagaaaag ttgaacgaga aacgtaaaat gatataaata tcaatatatt aaattagatt
      181 ttgcataaaa aacagactac ataatactgt aaaacacaac atatgcagtc actatgaatc
      241 aactacttag atggtattag tgacctgtag aattcgagct cgaccgacag ccttccaaat
      301 gttcttctca aacggaatcg tcgtatccag cctactcgct attgtcctca atgccgtatt
      361 aaatcataaa aagaaataag aaaaagaggt gcgagcctct tttttgtgtg acaaaataaa
      421 aacatctacc tattcatata cgctagtgtc atagtcctga aaatcatctg catcaagaac
      481 aatttcacaa ctcttatact tttctcttac aagtcgttcg gcttcatctg gattttcagc
      541 ctctatactt actaaacgtg ataaagtttc tgtaatttct actgtatcga cctgcagact
      601 ggctgtgtat aagggagcct gacatttata ttccccagaa catcaggtta atggcgtttt
      661 tgatgtcatt ttcgcggtgg ctgagatcag ccacttcttc cccgataacg gagaccggca
      721 cactggccat atcggtggtc atcatgcgcc agctttcatc cccgatatgc accaccgggt
      781 aaagttcacg ggagacttta tccgacagca gacgtgcact ggccaggggg atcaccatcc
      841 gtcgcccggg cgtgtcaata atatcactct gtacatccac aaacagatga taacggctct
      901 ctcttttata ggtgtaaacc ttaaactgca tttcaccagc ccctgttctc gtcagcaaaa
      961 gagccgttca tttcaataaa ccgggcgacc tcagccgtcc cttcctgatt ttccgctttc
     1021 cagcgttcgg cacgcagacg acgggcttca ttctgcatgg ttgtgcttac cagaccggag
     1081 atattgacat catatatgcc ttgagcaact gatagctgtc gctgtcaact gtcactgtaa
     1141 tacgctgctt catagcatac ctctttttga catacttcgg gtatacatat cagtatatat
     1201 tcttataccg caaaaatcag cgcgcaaata cgcatactgt tatctggctt ttagtaagcc
     1261 ggatccacgc ggcgtttacg ccccccctgc cactcatcgc agtactgttg taattcatta
     1321 agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc
     1381 atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag
     1441 ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag
     1501 acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac
     1561 gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag
     1621 agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc
     1681 catatcacca gctcaccgtc tttcattgcc atacggaatt ccggatgagc attcatcagg
     1741 cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt
     1801 aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga
     1861 aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg
     1921 atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg
     1981 cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaacg
     2041 tctcattttc gccaaaagtt ggcccagggc ttcccggtat caacagggac accaggattt
     2101 atttattctg cgaagtgatc ttccgtcaca ggtatttatt cggcgcaaag tgcgtcgggt
     2161 gatgctgcca acttagtcga cgctctagag ctccaaataa tgattttatt ttgactgata
     2221 gtgacctgtt cgttgcaaca aattgataag caatgctttt ttataatgcc aactttgtac
     2281 aaaaaagcag gctgaattcc cggggagctc acgcgttcgc gaatcgatcc gcggtctaga
     2341 aattcctggc attatcacat aatgaattat acattatata aagtaatgtg atttcttcga
     2401 agaatatact aaaaaatgag caggcaagat aaacgaaggc aaagatgaca gagcagaaag
     2461 ccctagtaaa gcgtattaca aatgaaacca agattcagat tgcgatctct ttaaagggtg
     2521 gtcccctagc gatagagcac tcgatcttcc cagaaaaaga ggcagaagca gtagcagaac
     2581 aggccacaca atcgcaagtg attaacgtcc acacaggtat agggtttctg gaccatatga
     2641 tacatgctct ggccaagcat tccggctggt cgctaatcgt tgagtgcatt ggtgacttac
     2701 acatagacga ccatcacacc actgaagact gcgggattgc tctcggtcaa gcttttaaag
     2761 aggccctagg ggccgtgcgt ggagtaaaaa ggtttggatc aggatttgcg cctttggatg
     2821 aggcactttc cagagcggtg gtagatcttt cgaacaggcc gtacgcagtt gtcgaacttg
     2881 gtttgcaaag ggagaaagta ggagatctct cttgcgagat gatcccgcat tttcttgaaa
     2941 gctttgcaga ggctagcaga attaccctcc acgttgattg tctgcgaggc aagaatgatc
     3001 atcaccgtag tgagagtgcg ttcaaggctc ttgcggttgc cataagagaa gccacctcgc
     3061 ccaatggtac caacgatgtt ccctccacca aaggtgttct tatgtagtga caccgattat
     3121 ttaaagctgc agcatacgat atatatacat gtgtatatat gtatacctat gaatgtcagt
     3181 aagtatgtat acgaacagta tgatactgaa gatgacaagg taatgcatca ttctatacgt
     3241 gtcattctga acgaggcgcg ctttcctttt ttctttttgc tttttctttt tttttctctt
     3301 gaactcgaga aaaaaaatat aaaagagatg gaggaacggg aaaaagttag ttgtggtgat
     3361 aggtggcaag tggtattccg taagaacaac aagaaaagca tttcatatta tggctgaact
     3421 gagcgaacaa gtgcaaaatt taagcatcaa cgacaacaac gagaatggtt atgttcctcc
     3481 tcacttaaga ggaaaaccaa gaagtgccag aaataacagt agcaactaca ataacaacaa
     3541 cggcggctac aacggtggcc gtggcggtgg cagcttcttt agcaacaacc gtcgtggtgg
     3601 ttacggcaac ggtggtttct tcggtggaaa caacggtggc agcagatcta acggccgttc
     3661 tggtggtaga tggatcgatg gcaaacatgt cccagctcca agaaacgaaa aggccgagat
     3721 cgccatattt ggtgtccccg agacccagct ttcttgtaca aagttgatat ctcgacccgg
     3781 gaattccgga ccggtacctg caggcgtacc agctttccct atagtgagtc gtattagagc
     3841 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     3901 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3961 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4021 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt
     4081 ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg
     4141 agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat
     4201 ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga
     4261 gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat
     4321 ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt
     4381 ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg
     4441 attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa
     4501 tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag
     4561 tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc
     4621 aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc
     4681 cagcggatag ttaatgatca gcccactgac ccgttgcgcg agaagattgt gcaccgccgc
     4741 tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg
     4801 atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga
     4861 ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg
     4921 aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg
     4981 gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac
     5041 atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta
     5101 tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tcaacgtaaa tgccgcttcg
     5161 ccttcgcgcg cgaattgcaa gctctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5221 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5281 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5341 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5401 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5461 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5521 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5581 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5641 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5701 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5761 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5821 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5881 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5941 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6001 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6061 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6121 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6181 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6241 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6301 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6361 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6421 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6481 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6541 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6601 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6661 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6721 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6781 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6841 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6901 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6961 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7021 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7081 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7141 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
     7201 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     7261 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     7321 gagtccacta ttaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     7381 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     7441 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     7501 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     7561 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     7621 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     7681 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     7741 ccagggtttt cccagtcacg acgtt
//
