LOCUS       sequence-8781-ex        4405 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 4405)
  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 4405)
  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: pB1H2wL-Prd
FEATURES             Location/Qualifiers
     source          1..4405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(88..93,94..111,112..118)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    126..142
                     /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     131..153
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     150..166
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     150..166
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             212..481
                     /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             491..514
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      1466..1921
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1553..1572)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1763..1784
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1947..2051
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2052..2120,2121..2912)
                     /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(2270..2289)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3083..3671
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3572..3591
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3825..3842
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3857..3997
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3923..3942)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4083..4105
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4099..4290)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gaattcaaaa aaatattgac aacataaaaa actttgtgtt atacttgtaa cgctacatgg
       61 agattaactc aatctagcta gagaggcttt acactttatg cttccggctc gtataatgtg
      121 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacggatt
      181 cactggaact ctaaaccaaa gagaggacac catggcacgc gtaactgttc aggacgctgt
      241 agagaaaatt ggtaaccgtt ttgacctggt actggtcgcc gcgcgtcgcg ctcgtcagat
      301 gcaggtaggc ggaaaggacc cgctcgtacc ggaagaaaac gataaaacca ctgtaatcgc
      361 gctgcgcgaa atcgaagaag gtctgatcaa caaccagatc ctcgacgttc gcgaacgcca
      421 ggaacagcaa gagcaggaag ccgctgaatt acaagccgtt accgctattg ctgaaggtcg
      481 tgcggccgcg gactacaagg atgacgacga caagttccgg accggttcca agacaccccc
      541 ccatggtacc aacagcggcc aggggcgcgt caatcaacta ggtggagttt tcatcaacgg
      601 tcgtcctttg cccaacaata ttcgtcttaa aatcgtcgag atggccgccg atggcattcg
      661 gccctgtgtg atctccagac agctacgtgt gtcccatggc tgcgtatcga agatcctgaa
      721 tcgctaccag gagactggct ccattagacc aggcgtaatt ggtggctcca agccgaggat
      781 agccacgccc gaaatcgaga accgaattga ggagtacaag cgcagtagcc cgggcatgtt
      841 ctcgtgggag atcagggaga agctgatccg cgagggtgtc tgcgacagga gcacagcacc
      901 atctgtgtcc gccatatcgc gcctggtgcg cggccgagat gctccattgg acaatgatat
      961 gtcttctgcc tctggatctc cggcgggtga tggcaccaaa gcatcgagtt cctgcggctc
     1021 cgatgtctca ggtggccatc acaacaacgg caagccctcc gatgaggaca tctctgactg
     1081 tgaaagtgag ccgggaatcg ccttgaagcg caaacagcgc cgctgcagga ccaccttttc
     1141 cgcttcccag ttggacgaac tggaacgcgc cttcgagcgc acccaatacc ctgatatcta
     1201 cacccgtgag gagctggccc agcgcaccaa tctcacggag gcacgcattc aggtgtggtt
     1261 cagcaatcgg cgtgctcgtc tccgcaagca gcacacctcg gtctcgggcg gagcacctgg
     1321 cggagcagct gcctcagtga gccattaatc tagagactag aaaaaggccg acaagtcccg
     1381 ctccgctgaa gatcctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     1441 ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     1501 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     1561 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     1621 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     1681 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     1741 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     1801 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     1861 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     1921 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     1981 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     2041 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     2101 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     2161 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     2221 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     2281 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     2341 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     2401 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     2461 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     2521 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     2581 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     2641 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     2701 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     2761 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     2821 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     2881 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     2941 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     3001 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     3061 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3121 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3181 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     3241 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3301 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3361 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3421 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3481 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3541 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3601 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3661 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3721 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3781 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3841 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3901 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     3961 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     4021 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     4081 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     4141 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     4201 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     4261 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
     4321 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
     4381 tgaacatgcc cggttactgg aacgg
//
