LOCUS       sequence-8780-ex        4499 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 4499)
  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 4499)
  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: pB1H2w5-Prd
FEATURES             Location/Qualifiers
     source          1..4499
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(12..17,18..35,36..42)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     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      1560..2015
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1647..1666)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1857..1878
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2041..2145
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2146..2214,2215..3006)
                     /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(2364..2383)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3177..3765
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3666..3685
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3919..3936
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3951..4091
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4017..4036)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4177..4199
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4193..4384)
                     /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 gctcgtataa 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 taccaacagc ggccaggggc
      661 gcgtcaatca actaggtgga gttttcatca acggtcgtcc tttgcccaac aatattcgtc
      721 ttaaaatcgt cgagatggcc gccgatggca ttcggccctg tgtgatctcc agacagctac
      781 gtgtgtccca tggctgcgta tcgaagatcc tgaatcgcta ccaggagact ggctccatta
      841 gaccaggcgt aattggtggc tccaagccga ggatagccac gcccgaaatc gagaaccgaa
      901 ttgaggagta caagcgcagt agcccgggca tgttctcgtg ggagatcagg gagaagctga
      961 tccgcgaggg tgtctgcgac aggagcacag caccatctgt gtccgccata tcgcgcctgg
     1021 tgcgcggccg agatgctcca ttggacaatg atatgtcttc tgcctctgga tctccggcgg
     1081 gtgatggcac caaagcatcg agttcctgcg gctccgatgt ctcaggtggc catcacaaca
     1141 acggcaagcc ctccgatgag gacatctctg actgtgaaag tgagccggga atcgccttga
     1201 agcgcaaaca gcgccgctgc aggaccacct tttccgcttc ccagttggac gaactggaac
     1261 gcgccttcga gcgcacccaa taccctgata tctacacccg tgaggagctg gcccagcgca
     1321 ccaatctcac ggaggcacgc attcaggtgt ggttcagcaa tcggcgtgct cgtctccgca
     1381 agcagcacac ctcggtctcg ggcggagcac ctggcggagc agctgcctca gtgagccatt
     1441 aatctagaga ctagaaaaag gccgacaagt cccgctccgc tgaagatcct ggcgtaatag
     1501 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     1561 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     1621 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     1681 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     1741 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     1801 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     1861 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     1921 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     1981 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     2041 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2101 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     2161 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     2221 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     2281 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     2341 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     2401 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     2461 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     2521 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     2581 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     2641 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     2701 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     2761 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     2821 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     2881 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     2941 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     3001 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     3061 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     3121 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3181 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3241 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3301 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     3361 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3421 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3481 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3541 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3601 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3661 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3721 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3781 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     3841 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     3901 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     3961 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     4021 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     4081 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     4141 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     4201 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     4261 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     4321 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     4381 tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac
     4441 gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgg
//
