LOCUS       sequence-381309-        3503 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 3503)
  AUTHORS   Wright DA, Thibodeau-Beganny S, Sander JD, Winfrey RJ, Hirsh AS, 
            Eichtinger M, Fu F, Porteus MH, Dobbs D, Voytas DF, Joung JK
  TITLE     Standardized reagents and protocols for engineering zinc finger 
            nucleases by modular assembly.
  JOURNAL   Nat Protoc. 2006;1(3):1637-52. doi: 10.1038/nprot.2006.259
  PUBMED    17406455
REFERENCE   2  (bases 1 to 3503)
  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."; date: "2006"; volume: "1(3):1637-52. doi"; pages: " 
            10.1038/nprot.2006.25"
COMMENT     Sequence Label: pGP-FF
FEATURES             Location/Qualifiers
     source          1..3503
                     /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      564..1019
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(651..670)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     861..882
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1045..1149
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1150..1218,1219..2010)
                     /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(1368..1387)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2181..2769
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2670..2689
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2923..2940
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2955..3095
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3021..3040)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3181..3203
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3197..3388)
                     /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 atctagagtg cagtgataac ctctactcac
      421 ctgcgtggat cctctgcaca gtagtaacta gagactagaa aaaggccgac aagtcccgct
      481 ccgctgaaga tcctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
      541 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
      601 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
      661 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
      721 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
      781 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
      841 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
      901 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
      961 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     1021 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     1081 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     1141 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     1201 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     1261 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     1321 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     1381 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     1441 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     1501 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     1561 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     1621 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     1681 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     1741 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     1801 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     1861 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     1921 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     1981 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     2041 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     2101 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     2161 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     2221 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2281 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     2341 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2401 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2461 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2521 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     2581 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     2641 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     2701 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     2761 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     2821 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     2881 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     2941 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3001 cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     3061 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     3121 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     3181 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     3241 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     3301 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     3361 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca
     3421 tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg
     3481 aacatgcccg gttactggaa cgg
//
