LOCUS       sequence-284-exp        6688 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 6688)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 6688)
  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: L4687
FEATURES             Location/Qualifiers
     source          1..6688
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2534..2554
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2576..2596
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2618..2638
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2660..2680
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2723..2751)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3499..3519
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4128..4150)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4250..4269
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4347..4365)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4433..4537
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4538..4606,4607..5398)
                     /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(4756..4775)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5569..6157
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6058..6077
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6311..6328
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6445..6466
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6481..6486,6487..6504,6505..6511)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6519..6535
                     /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     6524..6546
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6543..6559
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6543..6559
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgggc tgcaggtcgg ctataataag ttcttgaata aaataatttt cccgacaaaa
       61 catgagtatt tctttcgaaa ataaaagtgc aggctaatta gagattattc tgtaattaac
      121 tgcataattt gtcacgtgcc atagttttac attccactac gtcatagttc ttaaaatact
      181 aatctcctga aaatagaagt aggtgaagaa agtttaatta tcagttctaa aatgacaatt
      241 gatctttgga atatgttctg aaactaccga tcattgaaca gatgctattt gaatgatata
      301 gaattgtata tttgcaattt ctgaaacgcg ttcttaaagg cacacagatt aattcaaaag
      361 ggtctggccg caaaaaggtt tatggtggcc gattttgagt tttgtgtgtg attgcttttt
      421 cacaatcagt gttttcagga ttatgtgatg aactagatct tcaagtttcg ttacatttca
      481 tatgttttcg gaactcacga agtacatatt gggtattgtg ctcaaaaaat tcagcaatca
      541 gcttcgctcc gctgacttta gaacccaaaa aaatagtatg gccaaactga ctgtgttacg
      601 atcatttcaa tttttcaata catatttaag atttctaaga gtaagaaggt caaaaactgt
      661 tctggaatac atatatattt ttcaggttac aattagtcaa aaagtgcact gaaatatacg
      721 ttttaatttc acgaataacc caattagttc aatgtatttt tggtcaacca acgttaaagt
      781 ttggcttcca accaattatc atttctgatc aaccacaatg ttttttcttt atctgcaagt
      841 taatttttta tttttatcca gatgtttggc atatttttca attcttcact agcgcccact
      901 tcttgcactt ccggcgccct gaatctaatg catctgttgc aagaattgaa agaccaatca
      961 acacattgtt ttcttcacga gatactgaag aaaatgaata aaaacagaga aaaagagcca
     1021 tgtgattagt gacaactgtt gctaacagat accatagctt ggacttggta cgtgatggca
     1081 acgtatgggt caacaaaaat gattgcagag ggggtgcaaa acagtcaagt cgagaaaata
     1141 tgaaaaacag aaaacaaaga acagaaaaat gggtttgaga gtcagtataa tttataaaag
     1201 aaaaattgta catagaaatt aaccattttt gtagaagaag ttatttttca agcatcgtta
     1261 aaaattattc aaagcacctt atttcatatt taattttaaa catggttaaa tgaacaacac
     1321 ggtgcgcaat caggaaaact tgaaatctga aactgttgtt gtgatcttct tcgcaactgt
     1381 tcagatagca ctagtgtaat gttaagagtg cgcgaatata atggaatata atggatcaca
     1441 cctcctgcca tcaggtaaac gtctctgtta tcacatattt ccaactatta aatttttacc
     1501 ttttacagtt ttacattttt ttgaaaaaag taactttttg tcttcaaaat ccctgacgaa
     1561 aatatcaaat attttaatcg agactgcaga ggaaccgatt gatgatttgg aaaatccagc
     1621 tttacctgtg taagaactga aaagtttcat aaccctaggg tattcccagt tacattcccc
     1681 actggctaac aatagcaccc agtttttcat cacctttctt caaatttctc ggcgatttgt
     1741 taaaaacaaa atttgtgtcc cttctctgat atctctatgt ctctaaacac aagttcatcg
     1801 gaaaacgaag gagggtaggt gttggttggg ctcccgaagt gaaaatagaa gagcaagaat
     1861 agaatattag agagagagtg cagagagggc gggatagctc ccgggattcc gttttcttct
     1921 tctttatctt caacgatgat gtgtgtgcgt gttgtataga ttctgttgct cccccacaac
     1981 tcgctccgaa ggctcaatac aattcaattg atattggagg agagcctacc ggagtgggag
     2041 gataagaaga aacataagaa gaagaagaag aagaagcatg cttctggttt ttgatgctat
     2101 gaaaacggca caaaaagatg attgaggtcc cttttcaata ccttctctca tctttcaaat
     2161 cccattgaaa cctaaaactt ctcaccacgc tttaccattg ttctccaaaa acttatagca
     2221 atgtctataa cttttttatc tctgaaaagc agtgttccat ttttcttttt cctattttat
     2281 ttcaattgtt tctcacattt cgtttggatt ctttgcttgt caaccagctt cttcttccac
     2341 ttttaccgtc taattttcag ggcagggagc catcaaaccc acgaccacta gatccatcta
     2401 gaggatccta tcgattcgcg gccgctgtac acccccaaag gacccaaagg tatgtttcga
     2461 atgatactaa cataacatag aacattttca ggaggaccct tggagggtac cgagctcaga
     2521 aaaaatgact gctccaaaga agaagcgtaa ggtaccggca gaaaaaatga ctgctccaaa
     2581 gaagaagcgt aaggtaccgg cagaaaaaat gactgctcca aagaagaagc gtaaggtacc
     2641 ggcagaaaaa atgactgctc caaagaagaa gcgtaaggta ccggtagaaa aaatgagtaa
     2701 aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa
     2761 tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac
     2821 ccttaaattt atttgcacta ctggaaaact acctgttcca tgggtaagtt taaacatata
     2881 tatactaact aaccctgatt atttaaattt tcagccaaca cttgtcacta ctttcggtta
     2941 tggtcttcaa tgcttcgcca gatacccaga tcatatgaaa cagcatgact ttttcaagag
     3001 tgccatgccc gaaggttatg tacaggaaag aactatattt ttcaaagatg acgggaacta
     3061 caagacacgt aagtttaaac agttcggtac taactaacca tacatattta aattttcagg
     3121 tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa aaggtattga
     3181 ttttaaagaa gatggaaaca ttcttggaca caaattggaa tacaactata actcacacaa
     3241 tgtatacatc atggcagaca aacaaaagaa tggaatcaaa gttgtaagtt taaacatgat
     3301 tttactaact aactaatctg atttaaattt tcagaacttc aaaattagac acaacattga
     3361 agatggaagc gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc
     3421 tgtcctttta ccagacaacc attacctgtc ctaccaatct gccctttcga aagatcccaa
     3481 cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga ttacacatgg
     3541 catggatgaa ctatacaaat agcattcgta gaattgtaag tttaaacttt ctcatactaa
     3601 ctaacgcgct ctatttaaat tttcagaatt cgctagccgg ccatacaagt aatccggatg
     3661 atcgacgcca acgtcgttga attttcaaat tttaaatact gaatatttgt tttttttcct
     3721 attatttatt tattctcttt gtgttttttt tcttgctttc taaaaaatta attcaatcca
     3781 aatctaaaca tttttttttc tctttccgtc tcccaattcg tattccgctc ctctcatctg
     3841 aacacaatgt gcaagtttat ttatcttctc gctttcattt cattaggacg tggggggaat
     3901 tggtggaagg gggaaacaca caaaaggatg atggaaatga aataaggaca cacaatatgc
     3961 aacaacattc aattcagaaa tatggaggaa ggtttaaaag aaaacataaa aatatataga
     4021 ggaggaagga aaactagtaa aaaataagca aagaaattag gcgaacgatg agaattgtcc
     4081 tcgcttgggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     4141 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     4201 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     4261 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     4321 aaataccgca tcaggcggcc ttaagggcct cgtgatacgc ctatttttat aggttaatgt
     4381 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4441 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4501 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4561 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4621 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4681 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4741 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4801 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4861 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4921 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4981 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5041 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5101 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5161 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5221 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5281 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5341 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5401 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5461 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5521 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5581 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5641 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5701 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5761 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5821 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5881 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5941 gagataccta cagcgtgagc attgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6001 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6061 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6121 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6181 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6241 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     6301 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     6361 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     6421 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     6481 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     6541 cacaggaaac agctatgacc atgattacgc caagctgtaa gtttaaacat gatcttacta
     6601 actaactatt ctcatttaaa ttttcagagc ttaaaaatgg ctgaaatcac tcacaacgat
     6661 ggatacgcta acaacttgga aatgaaat
//
