LOCUS       sequence-9337-ex        7105 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 7105)
  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 7105)
  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: pGC204
FEATURES             Location/Qualifiers
     source          1..7105
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2885..2905
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2927..3604
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MVASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGTLIYKVKFRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFL"
     CDS             3647..4321
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGTLIYKVKFRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQA
                     LKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHHL
                     FL"
     promoter        4850..4954
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4955..5023,5024..5815)
                     /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(5173..5192)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5986..6574
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6475..6494
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6728..6745
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6862..6883
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6898..6903,6904..6921,6922..6928)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6936..6952
                     /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     6941..6963
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6960..6976
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6960..6976
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcaccggatg aagattctgt aatgtcattc acgtttggag atctgaaatt
       61 tgattacaaa acagaaccat ttattgatga atactacatt cgtgaatcga atcattctat
      121 atttgatgta ggtttcaaat aatatattca atcaatatca ttgtgtaaac acagtttgat
      181 gaagatgaaa cggtgaggat accacaaaag atgttgattt caagaccagg aatgcctcca
      241 ccacctccat ctcgtccatt aaatcttgct tcacctttga gaactctttt agagaaagaa
      301 cagaagtatg atgaagatcc agaaattcag ttaactccaa gacgattgaa ttcattatca
      361 gatattcagt aagttcctaa aaaattattc agccaagata tgtttcctgt accgaatagt
      421 ataaaaatat ttcagaattg gaaaactcca aaaacctttt aaaaagaaga agagtatgag
      481 ctttcttgaa atgttgttgt gtaatttcgg tgtaactcta aatcctgctg ctttcacaga
      541 cattttttgc ttatttcttc tcttacaaaa atgcagttta aactttaaaa gatttcaaaa
      601 aatatattgg acactcgcta tcacatttgc ccactgtata tccatgccag ctatttattt
      661 caatcaatga tttttccact ttgaaagtaa tattgtgcat ttccagggct cgtaaagtaa
      721 aacttggcgt ggatgaaaat cttcaacatg ctcgccttgt ctgtggtttg ttacctcaag
      781 attttgattc tccatcaact tccacgtaat gttttctcgc tgttaaaact caattgaatc
      841 acttcaattt tcaggtcgac gtcaatgata gattctggat atgaactttc aaaacgagat
      901 gcgtctacaa tggcttaatt atcacacttt taaaatattg ctactctaaa atgtcttttc
      961 aacttttcag tgtttatatc cggtttatat gtattacggg tcatgtcgtg tcaataaaat
     1021 ggtaattgta taagaagtgg gccaatagtg tgctctttta tttgtcatgt gtcctggtgg
     1081 cagaaataca gtaatttgat gcctacacgc aacagtaata ctttactaat ctcttttaac
     1141 taatatgtac acacaaactt tcatgacctt tgttcgttcg ttccctaatc ttagtaaaaa
     1201 agtatagaga agaggatatt gtgaatccat tttgtgtgtg taggtggaga aacgcacaca
     1261 caaatcagca aggaacaaga agttgagcaa aaaagattac ggtaagtgga caaacagata
     1321 acgcggccgt atttttggat ccaaaagcca aaattgttga tgaggaagat tgatgatgcg
     1381 caaattaggg agcagttttt ggaaggtcaa agagaagccc gtccgcacgc atctcccgcg
     1441 tacacaccac aaataaggta gtttactttt cggcggcatc cctttgcggc tgtacacaat
     1501 aatcttgtag aagatgattc taatacttgt aattggatta aggttagttg tgaatacgag
     1561 acagtgatac tgttcacaag ttttctttcg gggaaatgta tatgtatatg agtgaagtta
     1621 tgttgaaaaa ttgtcaaatt aagtcaaaga attcaatgta aaatttttaa gccaaactct
     1681 cagtgatatt aatagtctta atgtagacaa agcaagtgat acagttacat agttaagcac
     1741 gactatatgc gctttgtacc tactcaaagg gttactcaga aatcgctaga acctatcata
     1801 atgagagttt gttttttaaa tacgttagag caagataaat tcaagccaag ataatagatt
     1861 tgccttacag gcaactattt tctgactgat ttcctgaaat gtaaaaatta atagaaaaac
     1921 tatttaaagc aaagttttga gcaaaatcga atcctaaagt tactgaacta cgttcaagct
     1981 ttcggagcat taacctagaa taaatccagt gcgcgaaagt ttgggggatc attgcgaaca
     2041 cgaaatgagc ataaatatat tgggtgctca tttgccgaac cacttatagt tcgagcacta
     2101 cacttgatta ttgttagccg agatgctcat accccagcaa ataaataacg tgtgcaaaaa
     2161 gatagaattt gatctattat ggaaagttat gtattaagat atcgactttg gtacctaatc
     2221 gattattgtt attaatttgt gaaaggagcc ttgatgagat ttatctttaa ccattaattg
     2281 catacaatga ccactaattc aaatctattc cgcaattggg tccttgtctt tataaagaat
     2341 gccaaaatgt tgtactggtt tcaaaagtaa gtaggcatag tttcaagtgt aataaagttt
     2401 ttacataatc tatatatttg tcttgatgga aatatttaag tatccgatat aaaactaatt
     2461 acactagtct cagaagatct tttttttaat aacacaatct ttactagaaa tgaatttcaa
     2521 aaaaatagga ctaaattgta ctttccattc ctcttcactc tactcatcct cacatctccg
     2581 agatttgtca caaaagagat gaaattgtat actgagctta caagagtgcg cagagatatc
     2641 taagcataca aatctaccgt accgtcttcg ctttctccgc gatttgtggg ttggcgagcc
     2701 agagggagca caggtgggcg tgttgcaatc agcacaagag ggtcggttgt caattgcact
     2761 cgactctaga ggatccccgg gattggccaa aggacccaaa ggtatgtttc gaatgatact
     2821 aacataacat agaacatttt caggaggacc cttgagggta ccgagctcag aaaaaatgac
     2881 tgctccaaag aagaagcgta aggtaccggt agacgataag gatccgatgg tggcctcctc
     2941 cgaggacgtc atcaaagagt tcatgcgctt caaggtgcgc atggagggct ccgtgaacgg
     3001 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     3061 caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca
     3121 gttccagtac ggctccaagg cgtacgtgaa gcaccccgcc gacatccccg actacaagaa
     3181 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     3241 ggtgaccgtg acccaggact cctccctgca ggacggcacg ctgatctaca aggtgaagtt
     3301 ccgcggcacc aacttccccc ccgacggccc cgtaatgcag aagaagacca tgggctggga
     3361 ggcctccacc gagcgcctgt acccccgcga cggcgtgctg aagggcgaga tccaccaggc
     3421 cctgaagctg aaggacggcg gccactacct ggtggagttc aagaccatct acatggccaa
     3481 gaagcccgtg cagctgcccg gctactacta cgtggacacc aagctggaca tcacctccca
     3541 caacgaggac tacaccatcg tggaacagta cgagcgctcc gagggccgcc accacctgtt
     3601 cctggggcat ggcaccggca gcaccggcag cggcagctcc ggcaccgcct cctccgagga
     3661 cgtcatcaaa gagttcatgc gcttcaaggt gcgcatggag ggctccgtga acggccacga
     3721 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     3781 gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc cccagttcca
     3841 gtacggctcc aaggcgtacg tgaagcaccc cgccgacatc cccgactaca agaagctgtc
     3901 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     3961 cgtgacccag gactcctccc tgcaggacgg cacgctgatc tacaaggtga agttccgcgg
     4021 caccaacttc ccccccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     4081 caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccacc aggccctgaa
     4141 gctgaaggac ggcggccact acctggtgga gttcaagacc atctacatgg ccaagaagcc
     4201 cgtgcagctg cccggctact actacgtgga caccaagctg gacatcacct cccacaacga
     4261 ggactacacc atcgtggaac agtacgagcg ctccgagggc cgccaccacc tgttcctgta
     4321 ggaattcgct agccggccat acaagtaatc cggatgatcg acgccaacgt cgttgaattt
     4381 tcaaatttta aatactgaat atttgttttt tttcctatta tttatttatt ctctttgtgt
     4441 tttttttctt gctttctaaa aaattaattc aatccaaatc taaacatgag cggttttttt
     4501 tctctttccg tctcccaatt cgtattccgc tcctctcatc tgaacacaat gtgcaagttt
     4561 atttatcttc tcgctttcat ttcattagga cgtgggggga attggtggaa gggggaaaca
     4621 cacaaaagga tgatggaaat gaaataagga cacacaatat gcaacaacat tcaattcaga
     4681 aatatggagg aaggtttaaa agaaaacata aaaatatata gaggaggaag gaaaactagt
     4741 aaaaaataag caaagaaatt aggcgaacga tgagaattgt cctcgcttgg gcccaaaagg
     4801 cctttttttt ttggcgccga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4861 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4921 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4981 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     5041 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     5101 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     5161 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     5221 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     5281 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5341 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     5401 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5461 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     5521 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5581 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5641 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5701 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5761 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     5821 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5881 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5941 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6001 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6061 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6121 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6181 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6241 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6301 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6361 atacctacag cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6421 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6481 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6541 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6601 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6661 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     6721 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     6781 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     6841 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     6901 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     6961 aggaaacagc tatgaccatg attacgccaa gctgtaagtt taaacatgat cttactaact
     7021 aactattctc atttaaattt tcagagctta aaaatggctg aaatcactca caacgatgga
     7081 tacgctaaca acttggaaat gaaat
//
