LOCUS       sequence-103-exp        4536 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 4536)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 4536)
  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_70
FEATURES             Location/Qualifiers
     source          1..4536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             150..170
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(213..241)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     989..1009
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1976..1998)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2098..2117
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2195..2213)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2281..2385
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2386..2454,2455..3246)
                     /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(2604..2623)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3417..4005
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3906..3925
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4159..4176
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4293..4314
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4329..4334,4335..4352,4353..4359)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4367..4383
                     /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     4372..4394
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4391..4407
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4391..4407
                     /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 cttgcttgga
      121 gggtaccgag ctcagaaaaa atgactgctc caaagaagaa gcgtaaggta ccggtagaaa
      181 aaatgagtaa aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg
      241 gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg
      301 gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcca tgggtaagtt
      361 taaacatata tatactaact aaccctgatt atttaaattt tcagccaaca cttgtcacta
      421 ctttctgtta tggtgttcaa tgcttctcga gatacccaga tcatatgaaa cggcatgact
      481 ttttcaagag tgccatgccc gaaggttatg tacaggaaag aactatattt ttcaaagatg
      541 acgggaacta caagacacgt aagtttaaac agttcggtac taactaacca tacatattta
      601 aattttcagg tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa
      661 aaggtattga ttttaaagaa gatggaaaca ttcttggaca caaattggaa tacaactata
      721 actcacacaa tgtatacatc atggcagaca aacaaaagaa tggaatcaaa gttgtaagtt
      781 taaacatgat tttactaact aactaatctg atttaaattt tcagaacttc aaaattagac
      841 acaacattga agatggaagc gttcaactag cagaccatta tcaacaaaat actccaattg
      901 gcgatggccc tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcga
      961 aagatcccaa cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga
     1021 ttacacatgg catggatgaa ctatacaaat agcattcgta gaattccaac tgagcgccgg
     1081 tcgctaccat taccaacttg tctggtgtca aaaataatag gggccgctgt catcagagta
     1141 agtttaaact gagttctact aactaacgag taatatttaa attttcagca tctcgcgccc
     1201 gtgcctctga cttctaagtc caattactct tcaacatccc tacatgctct ttctccctgt
     1261 gctcccaccc cctatttttg ttattatcaa aaaacttctc ttaatttctt tgttttttag
     1321 cttcttttaa gtcacctcta acaatgaaat tgtgtagatt caaaaataga attaattcgt
     1381 aataaaaagt cgaaaaaaat tgtgctccct ccccccatta ataataattc tatcccaaaa
     1441 tctacacaat gttctgtgta cacttcttat gttttttact tctgataaat ttttttgaaa
     1501 catcatagaa aaaaccgcac acaaaatacc ttatcatatg ttacgtttca gtttatgacc
     1561 gcaattttta tttcttcgca cgtctgggcc tctcatgacg tcaaatcatg ctcatcgtga
     1621 aaaagttttg gagtattttt ggaatttttc aatcaagtga aagtttatga aattaatttt
     1681 cctgcttttg ctttttgggg tttcccctat tgtttgtcaa gatttcgagg acggcgtttt
     1741 tcttgctaaa atcacaagta ttgatgagca cgatgcaaga aagatcggaa gaaggtttgg
     1801 gtttgaggct cagtggaagg tgagtagaag ttgataattt gaaagtggag tagtgtctat
     1861 ggggtttttg ccttaaatga cagaatacat tcccaatata ccaaacataa ctgtttccta
     1921 ctagtcggcc gtacgggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     1981 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     2041 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     2101 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     2161 taaggagaaa ataccgcatc aggcggcctt aagggcctcg tgatacgcct atttttatag
     2221 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     2281 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2341 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     2401 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     2461 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     2521 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     2581 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     2641 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     2701 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     2761 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     2821 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     2881 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     2941 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     3001 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     3061 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     3121 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     3181 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     3241 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     3301 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     3361 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3421 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3481 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3541 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     3601 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3661 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3721 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3781 accgaactga gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga
     3841 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3901 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3961 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4021 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     4081 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     4141 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     4201 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     4261 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     4321 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     4381 caatttcaca caggaaacag ctatgaccat gattacgcca agctgtaagt ttaaacatga
     4441 tcttactaac taactattct catttaaatt ttcagagctt aaaaatggct gaaatcactc
     4501 acaacgatgg atacgctaac aacttggaaa tgaaat
//
