LOCUS       sequence-8785-ex        4055 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 4055)
  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 4055)
  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-mutOdd
FEATURES             Location/Qualifiers
     source          1..4055
                     /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      1116..1571
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1203..1222)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1413..1434
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1597..1701
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1702..1770,1771..2562)
                     /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(1920..1939)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2733..3321
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3222..3241
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3475..3492
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3507..3647
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3573..3592)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3733..3755
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3749..3940)
                     /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 ccccccatag atctagtcgt ccaaagaagc
      661 agttcatctg caagtactgc aaccgtcagt tcaccaagtc gtacaatctg ctcatccatg
      721 agagaaccca cacggacgag aggccttact cctgcgacat ctgcggcaag gccttccgtc
      781 gtcaggatca tctgcgtgac caccgttaca tccactccaa ggacaagccg ttcaagtgca
      841 gcgattgcgg caagggtttc tgccagtcgc gcaccctggc cgtgcacaag gtcacccatc
      901 tggaggaggg tccgcacaag tgtcccatct gccagcgcag cttcaaccag cgtgccaacc
      961 tcaagagtca cctccagagc cacagcgagc agagctaatc tagagactag aaaaaggccg
     1021 acaagtcccg ctccgctgaa gatcctggcg taatagcgaa gaggcccgca ccgatcgccc
     1081 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     1141 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     1201 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     1261 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     1321 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     1381 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     1441 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     1501 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     1561 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     1621 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     1681 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     1741 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     1801 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     1861 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     1921 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     1981 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     2041 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     2101 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     2161 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     2221 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     2281 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     2341 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     2401 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     2461 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     2521 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     2581 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     2641 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     2701 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2761 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2821 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     2881 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2941 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3001 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3061 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3121 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3181 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3241 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3301 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3361 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     3421 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     3481 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     3541 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     3601 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     3661 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     3721 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     3781 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
     3841 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
     3901 agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg
     3961 gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg
     4021 ttactgatga tgaacatgcc cggttactgg aacgg
//
