LOCUS       Exported                4273 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pGC76
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4273)
  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.
  PUBMED    18723890
REFERENCE   2  (bases 1 to 4273)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4273
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    complement(78..125)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             164..184
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             206..226
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             248..268
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             290..310
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(353..381)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1129..1149
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1713..1735)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1835..1854
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1932..1950)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2018..2122
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2123..2983
                     /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(2341..2360)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3154..3742
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3643..3662
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3896..3913
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4030..4051
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4066..4096
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4104..4120
                     /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     4109..4131
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4128..4144
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4128..4144
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ccgcggtttt ggcgcggatc cactagttct
       61 agcttcaaaa gcgctctgaa gttcctatac tttctagaga ataggaactt cggaatagga
      121 acttcaagat ccccgggtac cgagctcaga aaaaatgact gctccaaaga agaagcgtaa
      181 ggtaccggca gaaaaaatga ctgctccaaa gaagaagcgt aaggtaccgg cagaaaaaat
      241 gactgctcca aagaagaagc gtaaggtacc ggcagaaaaa atgactgctc caaagaagaa
      301 gcgtaaggta ccggtagaaa aaatgagtaa aggagaagaa cttttcactg gagttgtccc
      361 aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg
      421 tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta ctggaaaact
      481 acctgttcca tgggtaagtt taaacatata tatactaact aaccctgatt atttaaattt
      541 tcagccaaca cttgtcacta ctttctgtta tggtgttcaa tgcttctcga gatacccaga
      601 tcatatgaaa cggcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaaag
      661 aactatattt ttcaaagatg acgggaacta caagacacgt aagtttaaac agttcggtac
      721 taactaacca tacatattta aattttcagg tgctgaagtc aagtttgaag gtgataccct
      781 tgttaataga atcgagttaa aaggtattga ttttaaagaa gatggaaaca ttcttggaca
      841 caaattggaa tacaactata actcacacaa tgtatacatc atggcagaca aacaaaagaa
      901 tggaatcaaa gttgtaagtt taaacatgat tttactaact aactaatctg atttaaattt
      961 tcagaacttc aaaattagac acaacattga agatggaagc gttcaactag cagaccatta
     1021 tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc
     1081 cacacaatct gccctttcga aagatcccaa cgaaaagaga gaccacatgg tccttcttga
     1141 gtttgtaaca gctgctggga ttacacatgg catggacgaa ctatacaaaa gcccacaagc
     1201 tttggaattc gctagccggc catacaagta atccggatga tcgacgccaa cgtcgttgaa
     1261 ttttcaaatt ttaaatactg aatatttgtt ttttttccta ttatttattt attctctttg
     1321 tgtttttttt cttgctttct aaaaaattaa ttcaatccaa atctaaacat gagcggtttt
     1381 ttttctcttt ccgtctccca attcgtattc cgctcctctc atctgaacac aatgtgcaag
     1441 tttatttatc ttctcgcttt catttcatta ggacgtgggg ggaattggtg gaagggggaa
     1501 acacacaaaa ggatgatgga aatgaaataa ggacacacaa tatgcaacaa cattcaattc
     1561 agaaatatgg aggaaggttt aaaagaaaac ataaaaatat atagaggagg aaggaaaact
     1621 agtaaaaaat aagcaaagaa attaggcgaa cgatgagaat tgtcctcgct tgggcccttt
     1681 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     1741 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     1801 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     1861 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     1921 cggccttaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     1981 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2041 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2101 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2161 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2221 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     2281 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     2341 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     2401 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     2461 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     2521 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     2581 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     2641 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     2701 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     2761 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     2821 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     2881 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     2941 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3001 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3061 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3121 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3181 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3241 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3301 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3361 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3421 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3481 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3541 tgagcattga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3601 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3661 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3721 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3781 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3841 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     3901 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     3961 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     4021 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     4081 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     4141 tgaccatgat tacgccaagc tgtaagttta aacatgatct tactaactaa ctattctcat
     4201 ttaaattttc agagcttaaa aatggctgaa atcactcaca acgatggata cgctaacaac
     4261 ttggaaatga aat
//
