LOCUS       sequence-106027-        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"
     CDS             112..303
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(297..319)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    405..545
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     460..479
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(560..577)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(731..1319)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(811..830)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1490..2281,2282..2350))
                     /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     2113..2132
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2351..2455)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2481..2936)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2618..2639)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2830..2849
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(3114..3137)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3428..3450)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3439..3455
                     /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)."
     promoter        complement(join(3463..3469,3470..3487,3488..3493))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
ORIGIN
        1 tccagtaacc gggcatgttc atcatcagta acccgtatcg tgagcatcct ctctcgtttc
       61 atcggtatca ttacccccat gaacagaaat cccccttaca cggaggcatc agtgaccaaa
      121 caggaaaaaa ccgcccttaa catggcccgc tttatcagaa gccagacatt aacgcttctg
      181 gagaaactca acgagctgga cgcggatgaa caggcagaca tctgtgaatc gcttcacgac
      241 cacgctgatg agctttaccg cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc
      301 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
      361 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag
      421 tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac
      481 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
      541 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
      601 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
      661 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
      721 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
      781 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
      841 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
      901 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
      961 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1021 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     1081 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     1141 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     1201 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     1261 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     1321 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     1381 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     1441 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     1501 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     1561 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     1621 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     1681 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     1741 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     1801 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     1861 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     1921 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     1981 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     2041 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     2101 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     2161 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     2221 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     2281 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     2341 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     2401 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     2461 ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     2521 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     2581 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     2641 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     2701 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     2761 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     2821 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     2881 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca
     2941 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     3001 acgccaggat cttcagcgga gcgggacttg tcggcctttt tctagtctct agttactact
     3061 gtgcatgtct tcttacttag gatcctcact tagaagacct ccccgggtct agatttgtca
     3121 tcatcgtctt tgtagtcagc ggccgccaaa gcttggattt ttctcagggc cttttgcaca
     3181 atgtatttcc taataaaact ttgcttgaca ggttccgtca tcgtactacc caattgtgtt
     3241 ttttcaaaat tcttgcaggt ttcgatggct tcttgtaata agctgcgttg ctcctctgga
     3301 gaaaagattt ggttaatttg gttgaggacg tttggtacgg tattttgtga tgattgctgt
     3361 tgcaaagggt tgttgtccgc acttgaattg ggaggttgca tttgctgctg ttgaggcata
     3421 gctgtttcct gtgtgaaatt gttatccgct cacaagtcga cacattatac gagccggaag
     3481 cataaagtgt aaagcccgga attcccgt
//
