LOCUS       sequence-9310-ex        4389 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 4389)
  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 4389)
  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: pGC31
FEATURES             Location/Qualifiers
     source          1..4389
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        19..37
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    60..184
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(209..214,215..232,233..239)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             293..952
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(359..378)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1294..1599
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1478..1497
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1640..1764)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1780..1796)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1781..1797)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(1822..1840)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1850..1867)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1850..1866)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1859..1881)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2008..2463
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2095..2114)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2305..2326
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2489..2593
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2594..2662,2663..3454)
                     /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(2812..2831)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3625..4213
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4114..4133
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4367..4384
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aacctggctt atcgaaatta atacgactca ctatagggag accggcagat ctgatatcaa
       61 caagtttgta caaaaaagct gaacgagaaa cgtaaaatga tataaatatc aatatattaa
      121 attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat atccagtcat
      181 attggcggcc gcattaggca ccccaggctt tacactttat gcttccggct cgtataatgt
      241 gtggattttg agttaggatc cgtcgagatt ttcaggagct aaggaagcta aaatggagaa
      301 aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga
      361 ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc
      421 ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct
      481 tgcccgcctg atgaatgctc atccggaatt ccgtatggca atgaaagacg gtgagctggt
      541 gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc
      601 atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga
      661 tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt
      721 tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat
      781 ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt
      841 gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag
      901 aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt aaagatctgg
      961 atccggctta ctaaaagcca gataacagta tgcgtatttg cgcgctgatt tttgcggtat
     1021 aagaatatat actgatatgt atacccgaag tatgtcaaaa agaggtatgc tatgaagcag
     1081 cgtattacag tgacagttga cagcgacagc tatcagttgc tcaaggcata tatgatgtca
     1141 atatctccgg tctggtaagc acaaccatgc agaatgaagc ccgtcgtctg cgtgccgaac
     1201 gctggaaagc ggaaaatcag gaagggatgg ctgaggtcgc ccggtttatt gaaatgaacg
     1261 gctcttttgc tgacgagaac aggggctggt gaaatgcagt ttaaggttta cacctataaa
     1321 agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga cacgcccggg
     1381 cgacggatgg tgatccccct ggccagtgca cgtctgctgt cagataaagt ctcccgtgaa
     1441 ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac cgatatggcc
     1501 agtgtgccgg tctccgttat cggggaagaa gtggctgatc tcagccaccg cgaaaatgac
     1561 atcaaaaacg ccattaacct gatgttctgg ggaatataaa tgtcaggctc ccttatacac
     1621 agccagtctg caggtcgacc atagtgactg gatatgttgt gttttacagt attatgtagt
     1681 ctgtttttta tgcaaaatct aatttaatat attgatattt atatcatttt acgtttctcg
     1741 ttcagctttc ttgtacaaag tggttgatat caagcttatc gataccgtcg acctcgaggg
     1801 ggggcccggt acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg
     1861 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     1921 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     1981 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     2041 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2101 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2161 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2221 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2281 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2341 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     2401 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     2461 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     2521 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2581 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     2641 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     2701 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     2761 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     2821 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     2881 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     2941 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3001 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3061 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     3121 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     3181 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     3241 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     3301 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     3361 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     3421 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     3481 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     3541 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     3601 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3661 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3721 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     3781 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3841 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3901 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3961 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4021 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4081 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4141 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4201 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4261 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     4321 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     4381 cgaggaagc
//
