LOCUS       sequence-104-exp        4530 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 4530)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 4530)
  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_73
FEATURES             Location/Qualifiers
     source          1..4530
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             144..164
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(207..235)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     983..1003
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1970..1992)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2092..2111
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2189..2207)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2275..2379
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2380..2448,2449..3240)
                     /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(2598..2617)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3411..3999
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3900..3919
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4153..4170
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4287..4308
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4323..4328,4329..4346,4347..4353)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4361..4377
                     /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     4366..4388
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4385..4401
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4385..4401
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactagagg atccccggga ttggccaaag gacccaaagg
       61 tatgtttcga atgatactaa cataacatag aacattttca ggaggaccct tggagggtac
      121 cgagctcaga aaaaatgact gctccaaaga agaagcgtaa ggtaccggta gaaaaaatga
      181 gtaaaggaga agaacttttc actggagttg tcccaattct tgttgaatta gatggtgatg
      241 ttaatgggca caaattttct gtcagtggag agggtgaagg tgatgcaaca tacggaaaac
      301 ttacccttaa atttatttgc actactggaa aactacctgt tccatgggta agtttaaaca
      361 tatatatact aactaaccct gattatttaa attttcagcc aacacttgtc actactttct
      421 gttatggtgt tcaatgcttc tcgagatacc cagatcatat gaaacggcat gactttttca
      481 agagtgccat gcccgaaggt tatgtacagg aaagaactat atttttcaaa gatgacggga
      541 actacaagac acgtaagttt aaacagttcg gtactaacta accatacata tttaaatttt
      601 caggtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag ttaaaaggta
      661 ttgattttaa agaagatgga aacattcttg gacacaaatt ggaatacaac tataactcac
      721 acaatgtata catcatggca gacaaacaaa agaatggaat caaagttgta agtttaaaca
      781 tgattttact aactaactaa tctgatttaa attttcagaa cttcaaaatt agacacaaca
      841 ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg
      901 gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt tcgaaagatc
      961 ccaacgaaaa gagagaccac atggtccttc ttgagtttgt aacagctgct gggattacac
     1021 atggcatgga tgaactatac aaatagcatt cgtagaattc caactgagcg ccggtcgcta
     1081 ccattaccaa cttgtctggt gtcaaaaata ataggggccg ctgtcatcag agtaagttta
     1141 aactgagttc tactaactaa cgagtaatat ttaaattttc agcatctcgc gcccgtgcct
     1201 ctgacttcta agtccaatta ctcttcaaca tccctacatg ctctttctcc ctgtgctccc
     1261 accccctatt tttgttatta tcaaaaaact tctcttaatt tctttgtttt ttagcttctt
     1321 ttaagtcacc tctaacaatg aaattgtgta gattcaaaaa tagaattaat tcgtaataaa
     1381 aagtcgaaaa aaattgtgct ccctcccccc attaataata attctatccc aaaatctaca
     1441 caatgttctg tgtacacttc ttatgttttt tacttctgat aaattttttt gaaacatcat
     1501 agaaaaaacc gcacacaaaa taccttatca tatgttacgt ttcagtttat gaccgcaatt
     1561 tttatttctt cgcacgtctg ggcctctcat gacgtcaaat catgctcatc gtgaaaaagt
     1621 tttggagtat ttttggaatt tttcaatcaa gtgaaagttt atgaaattaa ttttcctgct
     1681 tttgcttttt ggggtttccc ctattgtttg tcaagatttc gaggacggcg tttttcttgc
     1741 taaaatcaca agtattgatg agcacgatgc aagaaagatc ggaagaaggt ttgggtttga
     1801 ggctcagtgg aaggtgagta gaagttgata atttgaaagt ggagtagtgt ctatggggtt
     1861 tttgccttaa atgacagaat acattcccaa tataccaaac ataactgttt cctactagtc
     1921 ggccgtacgg gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     1981 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     2041 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     2101 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     2161 gaaaataccg catcaggcgg ccttaagggc ctcgtgatac gcctattttt ataggttaat
     2221 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     2281 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2341 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2401 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2461 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2521 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     2581 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     2641 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2701 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2761 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2821 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2881 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     2941 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3001 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3061 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3121 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3181 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3241 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     3301 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3361 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3421 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3481 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3541 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     3601 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3661 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3721 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3781 ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3841 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3901 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3961 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4021 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     4081 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     4141 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     4201 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     4261 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     4321 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     4381 cacacaggaa acagctatga ccatgattac gccaagctgt aagtttaaac atgatcttac
     4441 taactaacta ttctcattta aattttcaga gcttaaaaat ggctgaaatc actcacaacg
     4501 atggatacgc taacaacttg gaaatgaaat
//
