LOCUS       sequence-51228-e        4907 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 4907)
  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 4907)
  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: pB1H2w5-12-C81S
FEATURES             Location/Qualifiers
     source          1..4907
                     /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      1968..2423
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2055..2074)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2265..2286
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2449..2553
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2554..2622,2623..3414)
                     /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(2772..2791)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3585..4173
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4074..4093
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4327..4344
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4359..4499
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4425..4444)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4585..4607
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4601..4792)
                     /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 ccggaccggc tccaagaccc cgccgcacgg cacggcgcgc ccatatgctt
      661 gccctgtcga gtcctgcgat cgccgctttt ctcgctcgga tgagcttacc cgccatatcc
      721 gcatccacac aggccagaag cccttccagt gtcgaatctg catgcgtaac ttcagtcgta
      781 gtgaccacct taccacccac atccgcaccc acaccggtac cgccattgaa ataaaagata
      841 aacagctcac aggattacgc tttattgacc tttttgcagg attaggtggc tttagacttg
      901 ctttagaatc ttgcggtgct gagtgcgttt attctaatga atgggataaa tatgcacaag
      961 aagtatatga gatgaatttt ggtgaaaagc ctgagggcga cattacccaa gtaaatgaga
     1021 aaaccattcc tgatcacgac attttatgtg cagggtttcc gtcccaagcg ttttctatta
     1081 gtggaaaaca aaaaggattc gaggacagca gaggtacgct cttttttgat attgcacgta
     1141 ttgtccgtga aaaaaaacct aaagtggttt ttatggaaaa tgtgaaaaat tttgcatcgc
     1201 atgataatgg aaatacgtta gaagttgtaa aaaatacaat gaatgaattg gactattctt
     1261 ttcatgctaa agtattaaat gctttagatt atgggattcc acagaaaagg gaacgtatct
     1321 atatgatttg ttttcgcaat gatctcaata ttcaaaattt ccaatttcca aaaccttttg
     1381 agcttaatac ttttgtgaaa gatttgttat tacctgatag cgaggtggaa cacttagtta
     1441 ttgatagaaa agatttggta atgacaaacc aagaaattga gcaaacaacc cccaaaacag
     1501 ttcgacttgg tattgtagga aaaggtgggc aaggagaacg aatttatagc acaagaggca
     1561 ttgcaattac cttatctgct tatggtggcg gcattttcgc taagacaggg ggatatttag
     1621 taaacgggaa gacacggaaa ttacacccta gagagtgtgc tagagtaatg ggctacccag
     1681 atagttataa agtccacccg tcaaccagcc aagcatataa acaatttggt aactcagttg
     1741 ttatcaatgt acttcaatat attgcttata acattggttc atcattaaat ttcaaaccat
     1801 atgggcccgg ggactacaag gatgacgacg acaactcgag ataaggatcc tctagagact
     1861 agaaaaaggc cgacaagtcc cgctccgctg aagatcctgg cgtaatagcg aagaggcccg
     1921 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
     1981 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2041 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2101 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2161 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2221 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2281 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     2341 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     2401 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     2461 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2521 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     2581 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     2641 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     2701 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     2761 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     2821 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     2881 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     2941 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     3001 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     3061 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     3121 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     3181 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     3241 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     3301 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     3361 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     3421 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     3481 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     3541 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3601 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3661 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3721 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3781 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     3841 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     3901 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     3961 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4021 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4081 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4141 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4201 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4261 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     4321 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     4381 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat
     4441 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
     4501 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     4561 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     4621 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
     4681 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     4741 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct
     4801 ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc
     4861 tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgg
//
