LOCUS       sequence-299-exp        5843 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 5843)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 5843)
  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"
COMMENT     Sequence Label: pJKL464
FEATURES             Location/Qualifiers
     source          1..5843
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1457..1477
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1520..1548)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2296..2316
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3283..3305)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3405..3424
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3502..3520)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3588..3692
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3693..3761,3762..4553)
                     /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(3911..3930)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4724..5312
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5213..5232
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5466..5483
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5600..5621
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5636..5641,5642..5659,5660..5666)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5674..5690
                     /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     5679..5701
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5698..5714
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5698..5714
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgaaa tcacaaattc catttttatg tcatacttca gatttaaaca aatttatttt
       61 atgtgtgttt ttttaatggt atgtcctaac gattttccta attgacaact attatagatt
      121 gaaaacacag aatgccaagt aacgtaagaa attttttttt cgaataccag acataatttc
      181 aataaaagaa aaatctttaa aaaaagttta actttatact acaataattt tggtttttat
      241 gaaggaatct gtattgcggc acatcatgta tttgctcaag gtttcgttgg ttgacatatt
      301 tttgtgatcc gtggaatgag cttgcataga gtttagtcat aattgggttt attttacata
      361 cagtttgaag tatgttttgt ctgttattcg aaatttttta gttagtttgg taagttgaaa
      421 agaatttaca aaatttaact aaacgaggac gctcatctcg attcttataa ttccataata
      481 attctacggt aaaagtcaag ttatgcctca aaacatgtaa tacataatta ccaaaactac
      541 ttaattaccg catttcctta gtattttaat agtggtgtag tcgaattttt tttattgctt
      601 tattagactc aaaattgtct gcaaacaccc aatttcataa tgaaacttat tgaaaacaat
      661 acactttgaa acaatttagt atttcaggaa taaggtcact ggaacttcga aaatgataat
      721 ttgaataacc gttaataaaa tattcaaacc aatttgcata atcttgattt tgtatcatga
      781 tggtatagaa tgggactttt gaaagatgtg aagtttcaat tcagtaagta caactttaaa
      841 attgggctgc aagatttctt cttaaatttt caacagtttc aaaacactga aatcatgctg
      901 aactatggtt agagactcga caactttcct aaattttgga gcagtttcaa tggttttcga
      961 atgtatacta catttgtacc attttatgtg ttagtgtagg ttgctttgct tctaccttca
     1021 ctctcaaatt cttttttccg cggtaatttt tcaactatcg aaacaaagaa gaagtcatga
     1081 ataactgtcc gcatagagtt ctcacccgtc ccctttcttt tcaccgagca gtcacgagag
     1141 aaagaagaag aagcggaccg ctgcagagat tcttcgtagc ggagccccgc ccatcctctg
     1201 ttacgcatat gtttggatct catttcttcc actcattaac gttccaggta tgtttttttt
     1261 gttgattttt taatgtgtta cttttcatat gtttattact tgaactgacc gatttcaaat
     1321 atgattttca cagatgtcta gaggatcccc gggattggcc aaaggaccca aaggtatgtt
     1381 tcgaatgata ctaacataac atagaacatt ttcaggagga cccttgaggg taccgagctc
     1441 agaaaaaatg actgctccaa agaagaagcg taaggtaccg gtagaaaaaa tgagtaaagg
     1501 agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg
     1561 gcacaaattt tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct
     1621 taaatttatt tgcactactg gaaaactacc tgttccatgg gtaagtttaa acatatatat
     1681 actaactaac cctgattatt taaattttca gccaacactt gtcactactt tctgttatgg
     1741 tgttcaatgc ttctcgagat acccagatca tatgaaacgg catgactttt tcaagagtgc
     1801 catgcccgaa ggttatgtac aggaaagaac tatatttttc aaagatgacg ggaactacaa
     1861 gacacgtaag tttaaacagt tcggtactaa ctaaccatac atatttaaat tttcaggtgc
     1921 tgaagtcaag tttgaaggtg atacccttgt taatagaatc gagttaaaag gtattgattt
     1981 taaagaagat ggaaacattc ttggacacaa attggaatac aactataact cacacaatgt
     2041 atacatcatg gcagacaaac aaaagaatgg aatcaaagtt gtaagtttaa acttggactt
     2101 actaactaac ggattatatt taaattttca gaacttcaaa attagacaca acattgaaga
     2161 tggaagcgtt caactagcag accattatca acaaaatact ccaattggcg atggccctgt
     2221 ccttttacca gacaaccatt acctgtccac acaatctgcc ctttcgaaag atcccaacga
     2281 aaagagagac cacatggtcc ttcttgagtt tgtaacagct gctgggatta cacatggcat
     2341 ggatgaacta tacaaatagc attcgtagaa ttccaactga gcgccggtcg ctaccattac
     2401 caacttgtct ggtgtcaaaa ataatagggg ccgctgtcat cagagtaagt ttaaactgag
     2461 ttctactaac taacgagtaa tatttaaatt ttcagcatct cgcgcccgtg cctctgactt
     2521 ctaagtccaa ttactcttca acatccctac atgctctttc tccctgtgct cccaccccct
     2581 atttttgtta ttatcaaaaa acttctctta atttctttgt tttttagctt cttttaagtc
     2641 acctctaaca atgaaattgt gtagattcaa aaatagaatt aattcgtaat aaaaagtcga
     2701 aaaaaattgt gctccctccc cccattaata ataattctat cccaaaatct acacaatgtt
     2761 ctgtgtacac ttcttatgtt ttttacttct gataaatttt tttgaaacat catagaaaaa
     2821 accgcacaca aaatacctta tcatatgtta cgtttcagtt tatgaccgca atttttattt
     2881 cttcgcacgt ctgggcctct catgacgtca aatcatgctc atcgtgaaaa agttttggag
     2941 tatttttgga atttttcaat caagtgaaag tttatgaaat taattttcct gcttttgctt
     3001 tttggggttt cccctattgt ttgtcaagat ttcgaggacg gcgtttttct tgctaaaatc
     3061 acaagtattg atgagcacga tgcaagaaag atcggaagaa ggtttgggtt tgaggctcag
     3121 tggaaggtga gtagaagttg ataatttgaa agtggagtag tgtctatggg gtttttgcct
     3181 taaatgacag aatacattcc caatatacca aacataactg tttcctacta gtcggccgta
     3241 cgggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     3301 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     3361 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     3421 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     3481 ccgcatcagg cggccttaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     3541 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3601 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3661 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     3721 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3781 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     3841 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     3901 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     3961 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     4021 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4081 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4141 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4201 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4261 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4321 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4381 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4441 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4501 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4561 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4621 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4681 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4741 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4801 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4861 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4921 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4981 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5041 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5101 acctacagcg tgagcattga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5161 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5221 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5281 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5341 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5401 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     5461 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     5521 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     5581 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     5641 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     5701 gaaacagcta tgaccatgat tacgccaagc tgtaagttta aacatgatct tactaactaa
     5761 ctattctcat ttaaattttc agagcttaaa aatggctgaa atcactcaca acgatggata
     5821 cgctaacaac ttggaaatga aat
//
