LOCUS       sequence-234-exp        4666 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 4666)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 4666)
  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: L4043
FEATURES             Location/Qualifiers
     source          1..4666
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             563..583
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             605..625
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             647..667
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             689..709
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(752..780)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1528..1548
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2106..2128)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2228..2247
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2325..2343)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2411..2515
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2516..2584,2585..3376)
                     /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(2734..2753)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3547..4135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4036..4055
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4289..4306
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4423..4444
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4459..4464,4465..4482,4483..4489)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4497..4513
                     /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     4502..4524
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4521..4537
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4521..4537
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ggatcaagag catttgaatc agaatatgga
       61 gaacggagca tgagcatttt cgaagttttt tagatgcact agaacaaagc gtgttggctt
      121 cctctgagcc cgctttcctt atatacccgc attctgcagc cttacagaat gttctagaag
      181 gtcctagatg cattcgtttg aaaatactcc cggtgggtgc aaagagacgc agacggaaaa
      241 tgtatctggg tctctttatt gtgtacacta cttttccatg taccgaatgt gagtcgccct
      301 ccttttgcaa caagcagctc gaatgttcta gaaaaaggtg gaaaatagta taaataccgt
      361 tgaaaataaa taccgaacaa catttgctct aattgtgaaa ttagaaatct tcaaactata
      421 atcatctcac tggatcctat cgattcgcgg ccgctgtaca cccccaaagg acccaaaggt
      481 atgtttcgaa tgatactaac ataacataga acattttcag gaggaccctt ggagggtacc
      541 gagctcagaa aaaatgactg ctccaaagaa gaagcgtaag gtaccggcag aaaaaatgac
      601 tgctccaaag aagaagcgta aggtaccggc agaaaaaatg actgctccaa agaagaagcg
      661 taaggtaccg gcagaaaaaa tgactgctcc aaagaagaag cgtaaggtac cggtagaaaa
      721 aatgagtaaa ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg
      781 tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg caacatacgg
      841 aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttccat gggtaagttt
      901 aaacatatat atactaacta accctgatta tttaaatttt cagccaacac ttgtcactac
      961 tttctgttat ggtgttcaat gcttctcgag atacccagat catatgaaac ggcatgactt
     1021 tttcaagagt gccatgcccg aaggttatgt acaggaaaga actatatttt tcaaagatga
     1081 cgggaactac aagacacgta agtttaaaca gttcggtact aactaaccat acatatttaa
     1141 attttcaggt gctgaagtca agtttgaagg tgataccctt gttaatagaa tcgagttaaa
     1201 aggtattgat tttaaagaag atggaaacat tcttggacac aaattggaat acaactataa
     1261 ctcacacaat gtatacatca tggcagacaa acaaaagaat ggaatcaaag ttgtaagttt
     1321 aaacatgatt ttactaacta actaatctga tttaaatttt cagaacttca aaattagaca
     1381 caacattgaa gatggaagcg ttcaactagc agaccattat caacaaaata ctccaattgg
     1441 cgatggccct gtccttttac cagacaacca ttacctgtcc acacaatctg ccctttcgaa
     1501 agatcccaac gaaaagagag accacatggt ccttcttgag tttgtaacag ctgctgggat
     1561 tacacatggc atggacgaac tatacaaaag cccacaagct ttggaattcg ctagccggcc
     1621 atacaagtaa tccggatgat cgacgccaac gtcgttgaat tttcaaattt taaatactga
     1681 atatttgttt tttttcctat tatttattta ttctctttgt gttttttttc ttgctttcta
     1741 aaaaattaat tcaatccaaa tctaaacatt tttttttctc tttccgtctc ccaattcgta
     1801 ttccgctcct ctcatctgaa cacaatgtgc aagtttattt atcttctcgc tttcatttca
     1861 ttaggacgtg gggggaattg gtggaagggg gaaacacaca aaaggatgat ggaaatgaaa
     1921 taaggacaca caatatgcaa caacattcaa ttcagaaata tggaggaagg tttaaaagaa
     1981 aacataaaaa tatatagagg aggaaggaaa actagtaaaa aataagcaaa gaaattaggc
     2041 gaacgatgag aattgtcctc gcttgggccc tttcgtctcg cgcgtttcgg tgatgacggt
     2101 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     2161 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     2221 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     2281 cacagatgcg taaggagaaa ataccgcatc aggcggcctt aagggcctcg tgatacgcct
     2341 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     2401 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2461 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     2521 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2581 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2641 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2701 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     2761 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     2821 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     2881 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     2941 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3001 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     3061 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3121 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3181 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3241 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3301 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3361 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3421 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3481 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3541 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3601 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3661 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     3721 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3781 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3841 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3901 aacgacctac accgaactga gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc
     3961 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4021 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4081 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4141 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4201 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4261 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     4321 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     4381 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     4441 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     4501 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agctgtaagt
     4561 ttaaacatga tcttactaac taactattct catttaaatt ttcagagctt aaaaatggct
     4621 gaaatcactc acaacgatgg atacgctaac aacttggaaa tgaaat
//
