LOCUS       Exported                4310 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pGC231
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4310)
  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 4310)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4310
                     /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             201..221
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             243..263
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             285..305
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             327..347
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(390..418)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1166..1186
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1750..1772)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1872..1891
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1969..1987)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2055..2159
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2160..3020
                     /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(2378..2397)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3191..3779
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3680..3699
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3933..3950
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4067..4088
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4103..4133
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4141..4157
                     /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     4146..4168
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4165..4181
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4165..4181
                     /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 ccccgggtat gtttcgaatg atactaacat aacatagaac attttcagga
      181 gctcagaaaa aatgactgct ccaaagaaga agcgtaaggt accggcagaa aaaatgactg
      241 ctccaaagaa gaagcgtaag gtaccggcag aaaaaatgac tgctccaaag aagaagcgta
      301 aggtaccggc agaaaaaatg actgctccaa agaagaagcg taaggtaccg gtagaaaaaa
      361 tgagtaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg
      421 atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca acatacggaa
      481 aacttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg gtaagtttaa
      541 acatatatat actaactaac cctgattatt taaattttca gccaacactt gtcactactt
      601 tctgttatgg tgttcaatgc ttctcgagat acccagatca tatgaaacgg catgactttt
      661 tcaagagtgc catgcccgaa ggttatgtac aggaaagaac tatatttttc aaagatgacg
      721 ggaactacaa gacacgtaag tttaaacagt tcggtactaa ctaaccatac atatttaaat
      781 tttcaggtgc tgaagtcaag tttgaaggtg atacccttgt taatagaatc gagttaaaag
      841 gtattgattt taaagaagat ggaaacattc ttggacacaa attggaatac aactataact
      901 cacacaatgt atacatcatg gcagacaaac aaaagaatgg aatcaaagtt gtaagtttaa
      961 acatgatttt actaactaac taatctgatt taaattttca gaacttcaaa attagacaca
     1021 acattgaaga tggaagcgtt caactagcag accattatca acaaaatact ccaattggcg
     1081 atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc ctttcgaaag
     1141 atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct gctgggatta
     1201 cacatggcat ggacgaacta tacaaaagcc cacaagcttt ggaattcgct agccggccat
     1261 acaagtaatc cggatgatcg acgccaacgt cgttgaattt tcaaatttta aatactgaat
     1321 atttgttttt tttcctatta tttatttatt ctctttgtgt tttttttctt gctttctaaa
     1381 aaattaattc aatccaaatc taaacatgag cggttttttt tctctttccg tctcccaatt
     1441 cgtattccgc tcctctcatc tgaacacaat gtgcaagttt atttatcttc tcgctttcat
     1501 ttcattagga cgtgggggga attggtggaa gggggaaaca cacaaaagga tgatggaaat
     1561 gaaataagga cacacaatat gcaacaacat tcaattcaga aatatggagg aaggtttaaa
     1621 agaaaacata aaaatatata gaggaggaag gaaaactagt aaaaaataag caaagaaatt
     1681 aggcgaacga tgagaattgt cctcgcttgg gccctttcgt ctcgcgcgtt tcggtgatga
     1741 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     1801 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     1861 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     1921 accgcacaga tgcgtaagga gaaaataccg catcaggcgg ccttaagggc ctcgtgatac
     1981 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     2041 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     2101 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     2161 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     2221 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     2281 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     2341 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     2401 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     2461 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     2521 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     2581 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     2641 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     2701 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     2761 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     2821 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     2881 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     2941 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3001 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     3061 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3121 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3181 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3241 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3301 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     3361 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3421 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3481 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3541 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc
     3601 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3661 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3721 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3781 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3841 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3901 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     3961 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     4021 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     4081 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     4141 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctgt
     4201 aagtttaaac atgatcttac taactaacta ttctcattta aattttcaga gcttaaaaat
     4261 ggctgaaatc actcacaacg atggatacgc taacaacttg gaaatgaaat
//
