LOCUS       sequence-101-exp        4532 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 4532)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 4532)
  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: pPD95_67
FEATURES             Location/Qualifiers
     source          1..4532
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             146..166
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(209..237)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     985..1005
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1972..1994)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2094..2113
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2191..2209)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2277..2381
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2382..2450,2451..3242)
                     /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(2600..2619)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3413..4001
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3902..3921
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4155..4172
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4289..4310
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4325..4330,4331..4348,4349..4355)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4363..4379
                     /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     4368..4390
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4387..4403
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4387..4403
                     /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 cttggagggt
      121 accgagctca gaaaaaatga ctgctccaaa gaagaagcgt aaggtaccgg tagaaaaaat
      181 gagtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga
      241 tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa
      301 acttaccctt aaatttattt gcactactgg aaaactacct gttccatggg taagtttaaa
      361 catatatata ctaactaacc ctgattattt aaattttcag ccaacacttg tcactacttt
      421 ctgttatggt gttcaatgct tctcgagata cccagatcat atgaaacggc atgacttttt
      481 caagagtgcc atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg
      541 gaactacaag acacgtaagt ttaaacagtt cggtactaac taaccataca tatttaaatt
      601 ttcaggtgct gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg agttaaaagg
      661 tattgatttt aaagaagatg gaaacattct tggacacaaa ttggaataca actataactc
      721 acacaatgta tacatcatgg cagacaaaca aaagaatgga atcaaagttg taagtttaaa
      781 catgatttta ctaactaact aatctgattt aaattttcag aacttcaaaa ttagacacaa
      841 cattgaagat ggaagcgttc aactagcaga ccattatcaa caaaatactc caattggcga
      901 tggccctgtc cttttaccag acaaccatta cctgtccaca caatctgccc tttcgaaaga
      961 tcccaacgaa aagagagacc acatggtcct tcttgagttt gtaacagctg ctgggattac
     1021 acatggcatg gatgaactat acaaatagca ttcgtagaat tccaactgag cgccggtcgc
     1081 taccattacc aacttgtctg gtgtcaaaaa taataggggc cgctgtcatc agagtaagtt
     1141 taaactgagt tctactaact aacgagtaat atttaaattt tcagcatctc gcgcccgtgc
     1201 ctctgacttc taagtccaat tactcttcaa catccctaca tgctctttct ccctgtgctc
     1261 ccacccccta tttttgttat tatcaaaaaa cttctcttaa tttctttgtt ttttagcttc
     1321 ttttaagtca cctctaacaa tgaaattgtg tagattcaaa aatagaatta attcgtaata
     1381 aaaagtcgaa aaaaattgtg ctccctcccc ccattaataa taattctatc ccaaaatcta
     1441 cacaatgttc tgtgtacact tcttatgttt tttacttctg ataaattttt ttgaaacatc
     1501 atagaaaaaa ccgcacacaa aataccttat catatgttac gtttcagttt atgaccgcaa
     1561 tttttatttc ttcgcacgtc tgggcctctc atgacgtcaa atcatgctca tcgtgaaaaa
     1621 gttttggagt atttttggaa tttttcaatc aagtgaaagt ttatgaaatt aattttcctg
     1681 cttttgcttt ttggggtttc ccctattgtt tgtcaagatt tcgaggacgg cgtttttctt
     1741 gctaaaatca caagtattga tgagcacgat gcaagaaaga tcggaagaag gtttgggttt
     1801 gaggctcagt ggaaggtgag tagaagttga taatttgaaa gtggagtagt gtctatgggg
     1861 tttttgcctt aaatgacaga atacattccc aatataccaa acataactgt ttcctactag
     1921 tcggccgtac gggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     1981 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     2041 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     2101 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     2161 gagaaaatac cgcatcaggc ggccttaagg gcctcgtgat acgcctattt ttataggtta
     2221 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     2281 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2341 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2401 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2461 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2521 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2581 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     2641 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     2701 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2761 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     2821 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2881 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     2941 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     3001 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3061 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3121 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3181 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3241 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     3301 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     3361 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3421 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3481 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3541 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3601 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3661 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3721 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3781 aactgagata cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg
     3841 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3901 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3961 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4021 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4081 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     4141 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     4201 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     4261 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     4321 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     4381 ttcacacagg aaacagctat gaccatgatt acgccaagct gtaagtttaa acatgatctt
     4441 actaactaac tattctcatt taaattttca gagcttaaaa atggctgaaa tcactcacaa
     4501 cgatggatac gctaacaact tggaaatgaa at
//
