LOCUS       sequence-171-exp        4755 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 4755)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1997
REFERENCE   2  (bases 1 to 4755)
  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: L3569
FEATURES             Location/Qualifiers
     source          1..4755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             369..389
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(432..460)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1208..1228
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2195..2217)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2317..2336
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2414..2432)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2500..2604
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2605..2673,2674..3465)
                     /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(2823..2842)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3636..4224
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4125..4144
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4378..4395
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4512..4533
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4548..4553,4554..4571,4572..4578)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4586..4602
                     /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     4591..4613
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4610..4626
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4610..4626
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ggatccccaa ttcttgagac ttccactatc
       61 ttctctggca ccctccacta cgcgtaggcg tctctcgctt cgtgtattcc cgggaagccg
      121 ggttcccgtc tctcccgccg ctgccgctgc cgcacacagc tttacacctc gtagaatccc
      181 caaagagggg cgtggcttgc gggtgccaac atcctcctgc cgaggaagaa gcaggcactc
      241 atcactcgca tcatcaacct cgggattggc caaaggaccc aaaggtatgt ttcgaatgat
      301 actaacataa catagaacat tttcaggagg acccttggag ggtaccgagc tcagaaaaaa
      361 tgactgctcc aaagaagaag cgtaaggtac cggtagaaaa aatgagtaaa ggagaagaac
      421 ttttcactgg agttgtccca attcttgttg aattagatgg tgatgttaat gggcacaaat
      481 tttctgtcag tggagagggt gaaggtgatg caacatacgg aaaacttacc cttaaattta
      541 tttgcactac tggaaaacta cctgttccat gggtaagttt aaacatatat atactaacta
      601 accctgatta tttaaatttt cagccaacac ttgtcactac tttcacttat ggtgttcaat
      661 gcttctcgag atacccagat catatgaaac agcatgactt tttcaagagt gccatgcccg
      721 aaggttatgt acaggaaaga actatatttt tcaaagatga cgggaactac aagacacgta
      781 agtttaaaca gttcggtact aactaaccat acatatttaa attttcaggt gctgaagtca
      841 agtttgaagg tgataccctt gttaatagaa tcgagttaaa aggtattgat tttaaagaag
      901 atggaaacat tcttggacac aaattggaat acaacttcaa ctcacacaac gtatacatca
      961 tggcagacaa acaaaagaat ggaatcaaag ttgtaagttt aaacatgatt ttactaacta
     1021 actaatctga tttaaatttt cagaacttca aaattagaca caacattgaa gatggaagcg
     1081 ttcaactagc agaccattat caacaaaata ctccaattgg cgatggccct gtccttttac
     1141 cagacaacca ttacctgtcc acacaatctg ccctttcgaa agatcccaac gaaaagagag
     1201 accacatggt ccttcttgag tttgtaacag ctgctgggat tacacatggc atggatgaac
     1261 tatacaaata gcattcgtag aattccaact gagcgccggt cgctaccatt accaacttgt
     1321 ctggtgtcaa aaataatagg ggccgctgtc atcagagtaa gtttaaactg agttctacta
     1381 actaacgagt aatatttaaa ttttcagcat ctcgcgcccg tgcctctgac ttctaagtcc
     1441 aattactctt caacatccct acatgctctt tctccctgtg ctcccacccc ctatttttgt
     1501 tattatcaaa aaacttctct taatttcttt gttttttagc ttcttttaag tcacctctaa
     1561 caatgaaatt gtgtagattc aaaaatagaa ttaattcgta ataaaaagtc gaaaaaaatt
     1621 gtgctccctc cccccattaa taataattct atcccaaaat ctacacaatg ttctgtgtac
     1681 acttcttatg ttttttactt ctgataaatt tttttgaaac atcatagaaa aaaccgcaca
     1741 caaaatacct tatcatatgt tacgtttcag tttatgaccg caatttttat ttcttcgcac
     1801 gtctgggcct ctcatgacgt caaatcatgc tcatcgtgaa aaagttttgg agtatttttg
     1861 gaatttttca atcaagtgaa agtttatgaa attaattttc ctgcttttgc tttttggggt
     1921 ttcccctatt gtttgtcaag atttcgagga cggcgttttt cttgctaaaa tcacaagtat
     1981 tgatgagcac gatgcaagaa agatcggaag aaggtttggg tttgaggctc agtggaaggt
     2041 gagtagaagt tgataatttg aaagtggagt agtgtctatg gggtttttgc cttaaatgac
     2101 agaatacatt cccaatatac caaacataac tgtttcctac tagtcggccg tacgggccct
     2161 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     2221 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     2281 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     2341 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     2401 ggcggcctta agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     2461 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     2521 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     2581 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     2641 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     2701 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     2761 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     2821 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     2881 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     2941 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     3001 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     3061 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     3121 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     3181 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     3241 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     3301 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     3361 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     3421 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     3481 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     3541 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     3601 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3661 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3721 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3781 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3841 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3901 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3961 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4021 cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4081 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4141 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4201 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4261 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     4321 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     4381 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     4441 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     4501 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     4561 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     4621 tatgaccatg attacgccaa gctgtaagtt taaacatgat cttactaact aactattctc
     4681 atttaaattt tcagagctta aaaatggctg aaatcactca caacgatgga tacgctaaca
     4741 acttggaaat gaaat
//
