LOCUS       sequence-224-exp        4712 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 4712)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 4712)
  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: L4018
FEATURES             Location/Qualifiers
     source          1..4712
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             145..165
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             187..207
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             229..249
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             271..291
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(334..362)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1110..1130
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2152..2174)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2274..2293
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2371..2389)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2457..2561
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2562..2630,2631..3422)
                     /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(2780..2799)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3593..4181
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4082..4101
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4335..4352
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4469..4490
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4505..4510,4511..4528,4529..4535)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4543..4559
                     /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     4548..4570
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4567..4583
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4567..4583
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ggatccccgg gattggccaa aggacccaaa
       61 ggtatgtttc gaatgatact aacataacat agaacatttt caggaggacc cttgagggta
      121 ccgagctcag aaaaaatgac tgctccaaag aagaagcgta aggtaccggc agaaaaaatg
      181 actgctccaa agaagaagcg taaggtaccg gcagaaaaaa tgactgctcc aaagaagaag
      241 cgtaaggtac cggcagaaaa aatgactgct ccaaagaaga agcgtaaggt accggtagaa
      301 aaaatgagta aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat
      361 ggtgatgtta atgggcacaa attttctgtc agtggagagg gtgaaggtga tgcaacatac
      421 ggaaaactta cccttaaatt tatttgcact actggaaaac tacctgttcc atgggtaagt
      481 ttaaacatat atatactaac taaccctgat tatttaaatt ttcagccaac acttgtcact
      541 actttctgtt atggtgttca atgcttctcg agatacccag atcatatgaa acggcatgac
      601 tttttcaaga gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat
      661 gacgggaact acaagacacg taagtttaaa cagttcggta ctaactaacc atacatattt
      721 aaattttcag gtgctgaagt caagtttgaa ggtgataccc ttgttaatag aatcgagtta
      781 aaaggtattg attttaaaga agatggaaac attcttggac acaaattgga atacaactat
      841 aactcacaca atgtatacat catggcagac aaacaaaaga atggaatcaa agttgtaagt
      901 ttaaacatga ttttactaac taactaatct gatttaaatt ttcagaactt caaaattaga
      961 cacaacattg aagatggaag cgttcaacta gcagaccatt atcaacaaaa tactccaatt
     1021 ggcgatggcc ctgtcctttt accagacaac cattacctgt ccacacaatc tgccctttcg
     1081 aaagatccca acgaaaagag agaccacatg gtccttcttg agtttgtaac agctgctggg
     1141 attacacatg gcatggatga actatacaaa tagcattcgt agaattgtaa gtttaaactt
     1201 tctcatacta actaacgcgc tctatttaaa ttttcagaat tccaactgag cgccggtcgc
     1261 taccattacc aacttgtctg gtgtcaaaaa taataggggc cgctgtcatc agagtaagtt
     1321 taaactgagt tctactaact aacgagtaat atttaaattt tcagcatctc gcgcccgtgc
     1381 ctctgacttc taagtccaat tactcttcaa catccctaca tgctctttct ccctgtgctc
     1441 ccacccccta tttttgttat tatcaaaaaa cttctcttaa tttctttgtt ttttagcttc
     1501 ttttaagtca cctctaacaa tgaaattgtg tagattcaaa aatagaatta attcgtaata
     1561 aaaagtcgaa aaaaattgtg ctccctcccc ccattaataa taattctatc ccaaaatcta
     1621 cacaatgttc tgtgtacact tcttatgttt tttacttctg ataaattttt ttgaaacatc
     1681 atagaaaaaa ccgcacacaa aataccttat catatgttac gtttcagttt atgaccgcaa
     1741 tttttatttc ttcgcacgtc tgggcctctc atgacgtcaa atcatgctca tcgtgaaaaa
     1801 gttttggagt atttttggaa tttttcaatc aagtgaaagt ttatgaaatt aattttcctg
     1861 cttttgcttt ttggggtttc ccctattgtt tgtcaagatt tcgaggacgg cgtttttctt
     1921 gctaaaatca caagtattga tgagcacgat gcaagaaaga tcggaagaag gtttgggttt
     1981 gaggctcagt ggaaggtgag tagaagttga taatttgaaa gtggagtagt gtctatgggg
     2041 tttttgcctt aaatgacaga atacattccc aatataccaa acataactgt ttcctactag
     2101 tcggccgtac gggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     2161 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     2221 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     2281 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     2341 gagaaaatac cgcatcaggc ggccttaagg gcctcgtgat acgcctattt ttataggtta
     2401 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     2461 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2521 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2581 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2641 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2701 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2761 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     2821 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     2881 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2941 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     3001 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     3061 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     3121 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     3181 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3241 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3301 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3361 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3421 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     3481 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     3541 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3601 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3661 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3721 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3781 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3841 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3901 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3961 aactgagata cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg
     4021 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4081 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4141 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4201 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4261 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     4321 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     4381 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     4441 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     4501 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     4561 ttcacacagg aaacagctat gaccatgatt acgccaagct gtaagtttaa acatgatctt
     4621 actaactaac tattctcatt taaattttca gagcttaaaa atggctgaaa tcactcacaa
     4681 cgatggatac gctaacaact tggaaatgaa at
//
