LOCUS       sequence-8786-ex        3966 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 3966)
  AUTHORS   Noyes MB, Meng X, Wakabayashi A, Sinha S, Brodsky MH, Wolfe SA
  TITLE     A systematic characterization of factors that regulate Drosophila 
            segmentation via a bacterial one-hybrid system.
  JOURNAL   Nucleic Acids Res. 2008 May;36(8):2547-60. doi: 10.1093/nar/gkn048. 
            Epub 2008 Mar 10.
  PUBMED    18332042
REFERENCE   2  (bases 1 to 3966)
  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: 
            "10.1093/nar/gkn048"; journalName: "Nucleic Acids Res"; date: 
            "2008-05"; volume: "36"; issue: "8"; pages: "2547-60"
COMMENT     Sequence Label: pB1H2w2-zif268
FEATURES             Location/Qualifiers
     source          1..3966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    50..66
                     /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     55..77
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             306..575
                     /codon_start=1
                     /gene="E. coli rpoZ"
                     /product="DNA-directed RNA polymerase subunit omega"
                     /label=RNAP omega
                     /note="promotes RNA polymerase assembly"
                     /translation="MARVTVQDAVEKIGNRFDLVLVAARRARQMQVGGKDPLVPEENDK
                     TTVIALREIEEGLINNQILDVRERQEQQEQEAAELQAVTAIAEGR"
     CDS             585..608
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      1027..1482
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1114..1133)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1324..1345
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1508..1612
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1613..1681,1682..2473)
                     /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     complement(1831..1850)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2644..3232
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3133..3152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3386..3403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3418..3558
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3484..3503)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3644..3666
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3660..3851)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gaattccggg ctttacactt tatgcttccg gctcgcatta tgtgtcgact tgtgagcgga
       61 taacaatttc acacaggaaa cagctatgcc tcaacagcag caaatgcaac ctcccaattc
      121 aagtgcggac aacaaccctt tgcaacagca atcatcacaa aataccgtac caaacgtcct
      181 caaccaaatt aaccaaatct tttctccaga ggagcaacgc agcttattac aagaagccat
      241 cgaaacctgc aagaattttg aaaaaacaca attgtaagtc tcggctccaa gaaggagata
      301 tacccatggc acgcgtaact gttcaggacg ctgtagagaa aattggtaac cgttttgacc
      361 tggtactggt cgccgcgcgt cgcgctcgtc agatgcaggt aggcggaaag gacccgctcg
      421 taccggaaga aaacgataaa accactgtaa tcgcgctgcg cgaaatcgaa gaaggtctga
      481 tcaacaacca gatcctcgac gttcgcgaac gccaggaaca gcaagagcag gaagccgctg
      541 aattacaagc cgttaccgct attgctgaag gtcgtgcggc cgcggactac aaggatgacg
      601 acgacaagtt ccggaccggt tccaagacac ccccccatgg tacccgccca tatgcttgcc
      661 ctgtcgagtc ctgcgatcgc cgcttttctc gctcggatga gcttacccgc catatccgca
      721 tccataccgg tcagaagccc ttccagtgtc gaatctgcat gcgtaacttc agtcgtagtg
      781 accaccttac cacccacatc cgcacccaca caggcgagaa gccttttgcc tgtgacattt
      841 gtgggaggaa gtttgccagg agtgatgaac gcaagaggca taccaaaatc catttaagac
      901 agaaggacgt ctagagacta gaaaaaggcc gacaagtccc gctccgctga agatcctggc
      961 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     1021 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     1081 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     1141 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     1201 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     1261 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     1321 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     1381 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     1441 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     1501 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     1561 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     1621 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     1681 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     1741 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     1801 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     1861 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     1921 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     1981 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     2041 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     2101 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     2161 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     2221 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     2281 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     2341 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     2401 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     2461 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     2521 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     2581 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2641 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2701 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2761 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     2821 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2881 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2941 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3001 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3061 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     3361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     3421 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
     3481 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     3541 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     3601 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     3661 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     3721 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     3781 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     3841 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
     3901 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
     3961 gaacgg
//
