LOCUS       sequence-9245-ex        6774 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 6774)
  AUTHORS   Voutev R, Hubbard EJ
  TITLE     A ""FLP-Out"" system for controlled gene expression in 
            Caenorhabditis elegans.
  JOURNAL   Genetics. 2008 Sep;180(1):103-19. doi: 10.1534/genetics.108.090274.
  PUBMED    18723890
REFERENCE   2  (bases 1 to 6774)
  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: 
            "10.1534/genetics.108"; journalName: "Genetics"; date: "2008-09"; 
            volume: "180"; issue: "1"; pages: "103-19"
COMMENT     Sequence Label: pGC133
FEATURES             Location/Qualifiers
     source          1..6774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             508..1779
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=FLP
                     /note="FLP is a site-specific recombinase from 
                     Saccharomyces cerevisiae. Recombination occurs at FRT 
                     sequences."
                     /translation="MPQFDILCKTPPKVLVRQFVERFERPSGEKIALCAAELTYLCWMI
                     THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI
                     IPYYGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN
                     SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET
                     KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR
                     SYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR
                     TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR
                     YPAWNGIISQEVLDYLSSYINRRI"
     CDS             complement(2479..2490)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        4525..4629
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4630..4698,4699..5490)
                     /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(4848..4867)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5661..6249
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6150..6169
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6403..6420
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6537..6558
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6573..6578,6579..6596,6597..6603)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6611..6627
                     /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     6616..6638
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6635..6651
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6635..6651
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ggatcaccaa aaacggaacg ttgagctgga
       61 cggaaatagt ggtaaagtga catgattata gtttgaagat ttctaatttc acaattagag
      121 caaatgttgt tcggtattta ttttcaacgg tatttatact attttccacc tttttctaga
      181 acattcgagc tgcttgttgc aaaaggaggg cgactcacat tcggtacatg gaaaagtagt
      241 gtacacaata aagagaccca gatacatttt ccgtctgcgt ctctttgcac ccaccgggag
      301 tattttcaaa cgaatgcatc taggaccttc tagaacattc tgtaaggctg cagaatgcgg
      361 gtatataagg aaagcgggct cagaggaagc caacacgctt tgttctagtg catctaaaaa
      421 acttcgaaaa tcctcatcgg gatcctatcg attcgcggcc gctgtacacc cgggtgccct
      481 atagtgagtc gtattggtac caaaaacatg ccacaatttg atatattatg taaaacacca
      541 cctaaggtgc ttgttcgtca gtttgtggaa aggtttgaaa gaccttcagg tgagaaaata
      601 gcattatgtg ctgctgaact aacctattta tgttggatga ttacacataa cggaacagca
      661 atcaagagag ccacattcat gagctataat actatcataa gcaattcgct gagtttcgat
      721 attgtcaata aatcactcca gtttaaatac aagacgcaaa aagcaacaat tctggaagcc
      781 tcattaaaga aattgattcc tgcttgggaa tttacaatta ttccttacta tggacaaaaa
      841 catcaatctg atatcactga tattgtaagt agtttgcaat tacagttcga atcatcggaa
      901 gaagcagata agggaaatag ccacagtaaa aaaatgctta aagcacttct aagtgagggt
      961 gaaagcatct gggagatcac tgagaaaata ctaaattcgt ttgagtatac ttcgagattt
     1021 acaaaaacaa aaactttata ccaattcctc ttcctagcta ctttcatcaa ttgtggaaga
     1081 ttcagcgata ttaagaacgt tgatccgaaa tcatttaaat tagtccaaaa taagtatctg
     1141 ggagtaataa tccagtgttt agtgacagag acaaagacaa gcgttagtag gcacatatac
     1201 ttctttagcg caaggggtag gatcgatcca cttgtatatt tggatgaatt tttgaggaat
     1261 tctgaaccag tcctaaaacg agtaaatagg accggcaatt cttcaagcaa taaacaggaa
     1321 taccaattat taaaagataa cttagtcaga tcgtacaata aagctttgaa gaaaaatgcg
     1381 ccttattcaa tctttgctat aaaaaatggc ccaaaatctc acattggaag acatttgatg
     1441 acctcatttc tttcaatgaa gggcctaacg gagttgacta atgttgtggg aaattggagc
     1501 gataagcgtg cttctgccgt ggccaggaca acgtatactc atcagataac agcaatacct
     1561 gatcactact tcgcactagt ttctcggtac tatgcatatg atccaatatc aaaggaaatg
     1621 atagcattga aggatgagac taatccaatt gaggagtggc agcatataga acagctaaag
     1681 ggtagtgctg aaggaagcat acgatacccc gcatggaatg ggataatatc acaggaggta
     1741 ctagactacc tttcatccta cataaataga cgcatataag tacgcattta agcataaaca
     1801 cgcactatgc cgttcttctc atgtatatat atatacaggc aacacgcaga tataggtgcg
     1861 acgtgaacag tgagctgtat gtgcggggat ccactagttc tagccggcca tacaagtaat
     1921 ccggatgatc gacgccaacg tcgttgaatt ttcaaatttt aaatactgaa tatttgtttt
     1981 ttttcctatt atttatttat tctctttgtg ttttttttct tgctttctaa aaaattaatt
     2041 caatccaaat ctaaacatga gcggtttttt ttctctttcc gtctcccaat tcgtattccg
     2101 ctcctctcat ctgaacacaa tgtgcaagtt tatttatctt ctcgctttca tttcattagg
     2161 acgtgggggg aattggtgga agggggaaac acacaaaagg atgatggaaa tgaaataagg
     2221 acacacaata tgcaacaaca ttcaattcag aaatatggag gaaggtttaa aagaaaacat
     2281 aaaaatatat agaggaggaa ggaaaactag tctagaatcc tatgcttgca ctttgagcca
     2341 atttatccaa gtccttgtaa aaagtattcg aaaattgtta acggataaaa tgtttattat
     2401 aatatcaaaa acaatttgtc agttgaccac tttttgatat atttgacaga aaccgggatg
     2461 aattggctca aagttagggc gcccttctat tacagggttt ctgataacaa accggttatt
     2521 aactcccaac aagggatgat ttcaattcat catgctcaaa ttgacccaaa ttaagttaca
     2581 tgacaaattc atcgcccttt tcaactcttc ttggctcatc atctgttatt ctgttcctat
     2641 tctgtctgca ccctataccc ctttgcatac tctctcgtca ttcctttctt ggatagtgtc
     2701 ttcttctccc cagctctgct acttctatga cttgcgcccg ctgcttttcc gctcgttcct
     2761 ctcctctctg acgtcttctt ctgcttcttt tctccttcct ttttcccttc tcaatcttcc
     2821 tctttttccc atttctgtca atcattcgaa gaagaagaag aagaagaccc tcattcattt
     2881 catttttttt tctgtcggtg tgtgctgctc ggttaagagt gagctctcta ttccacgtct
     2941 tcttcttttt cttcttcgat tcgaatcaat cactccacca aaacgcattc gtttttggga
     3001 ttcaccccgc ggttcgcaac aggtttcttt ttcaaattat tagcgttata aatagaaaaa
     3061 tggtggagtt tcaataaaaa tgataattta caaagtgtat tttgattaca tgtactcaaa
     3121 aaggttgaaa tttttcatac cagttttccg gaaatccatc tgatatcatt atcgtatttt
     3181 tctttttaaa aatgttttca aaaaaaaaca aaatatagct ggttatttgg caccctctaa
     3241 ttaccatttt cctgtcacac cacactcttt tccttcttcc acttctttcc gttttcagcc
     3301 gcttccaacc aaaccgatat gaaagccgag caacaacaat cgattccacc cggctcggcg
     3361 accttcccgt cgcaggtgag actcagaaaa ctagagaaac cgctcaacta actcttgata
     3421 tccgatttca ttcttttctc ttttcttttt gttgaacttt tccatatttc cagatgccac
     3481 ggccaccacc aagcaccgaa caaggaatca caacggaatc ggagcttgcg aagaaagctc
     3541 aaatcactcc gaacgacgtt ttagcacttc cgggaatcac tcaaggtatc ttgctccctt
     3601 tggatttcag tttaaaacat aaatttagga ttcttatgct ccccatctgc gaacatctat
     3661 aacatcgagt tcaccaagtt ccaaatccgt gatctggaca ctgagcaagt gcttttcgag
     3721 atcgccaaac cggagaacga tcaggagaat gatgagtcgc cacaggagtc ggcaagatac
     3781 gtgcgttata gatttgctcc aaactttttg aaactcaaaa cggtcggagc aactgtggaa
     3841 ttcaaagtag gagacatccc aatccatcat ttccgaatga tcgaacgtca cttcttcaaa
     3901 gatcgccttc tgaagtgttt tgactttgaa ttcggattct gtattccgaa ttcacgaaac
     3961 aactgtgaac atatctatga gttccctcaa ctctctcaac aactcagtga gtcattattc
     4021 taaaaagtac aattcaaaag actaatctct ttttcagtgg acgacatgat caacaatcca
     4081 aacgagactc gttctgacag cttctatttc gtcgataaca aactcgtcat gcacaacaaa
     4141 gccgactact catatgatgc ataaatattt aatacaaaaa tgttctggat aattattctg
     4201 tcgaatagaa aaaaaactcc aaatgtgatt aaattccaat aattcctgtc tagtttgttc
     4261 cttccccttc cccttctcat gttcaatgca ttcctaagct tttcagttcc ccccttgttt
     4321 ctatattttt tcgctgtcct gtcacactcg ctaaaaacac taatcacacg gaaatctgtt
     4381 ttcaataaaa actccaactt taactcattt tcaatttcaa ctgaaagatt ttttcattag
     4441 agaatgtcta ggcatgctct agagggccca aaaggccttt tttttttggc gccgacgtca
     4501 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4561 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4621 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4681 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4741 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4801 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4861 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4921 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4981 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5041 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5101 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5161 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5221 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5281 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5341 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5401 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5461 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5521 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5581 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5641 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5701 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5761 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     5821 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5881 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5941 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6001 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gcattgagaa
     6061 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6121 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6181 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6241 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6301 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6361 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     6421 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     6481 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     6541 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     6601 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     6661 gccaagctgt aagtttaaac atgatcttac taactaacta ttctcattta aattttcaga
     6721 gcttaaaaat ggctgaaatc actcacaacg atggatacgc taacaacttg gaaa
//
