LOCUS       sequence-10054-e        3508 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 3508)
  AUTHORS   Maeder ML, Thibodeau-Beganny S, Sander JD, Voytas DF, Joung JK
  TITLE     Oligomerized pool engineering (OPEN): an 'open-source' protocol for 
            making customized zinc-finger arrays.
  JOURNAL   Nat Protoc. 2009. 4(10):1471-501.
  PUBMED    19798082
REFERENCE   2  (bases 1 to 3508)
  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"; journalName: "Nat Protoc.
            2009. 4(10):1471-501."
COMMENT     Sequence Label: pMLM36 (aka pBR-UV5-GP-FD2)
FEATURES             Location/Qualifiers
     source          1..3508
                     /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             368..391
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      569..1024
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(656..675)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     866..887
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1050..1154
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1155..1223,1224..2015)
                     /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(1373..1392)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2186..2774
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2675..2694
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2928..2945
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2960..3100
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3026..3045)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3186..3208
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3202..3393)
                     /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 attgggtagt acgatgacgg aacctgtcaa
      301 gcaaagtttt attaggaaat acattgtgca aaaggccctg agaaaaatcc aagctttggc
      361 ggccgctgac tacaaagacg atgatgacaa atctagaccc ggggaggtct tctaagtgag
      421 gatcctaagt aagaagacat gcacagtagt aactagagac tagaaaaagg ccgacaagtc
      481 ccgctccgct gaagatcctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
      541 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
      601 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
      661 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
      721 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
      781 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
      841 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
      901 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
      961 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     1021 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1081 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     1141 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1201 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1261 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1321 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     1381 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     1441 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     1501 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     1561 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     1621 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     1681 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     1741 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     1801 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     1861 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     1921 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     1981 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     2041 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     2101 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2161 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2221 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2281 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     2341 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2401 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     2461 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     2521 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     2581 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     2641 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     2701 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     2761 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     2821 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     2881 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     2941 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3001 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     3061 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc
     3121 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     3181 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     3241 gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc
     3301 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
     3361 catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct
     3421 gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga
     3481 tgatgaacat gcccggttac tggaacgg
//
