LOCUS       sequence-225-exp        4717 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 4717)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 4717)
  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: L4019
FEATURES             Location/Qualifiers
     source          1..4717
                     /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"
     CDS             192..212
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             234..254
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             276..296
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(339..367)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1115..1135
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2157..2179)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2279..2298
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2376..2394)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2462..2566
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2567..2635,2636..3427)
                     /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(2785..2804)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3598..4186
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4087..4106
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4340..4357
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4474..4495
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4510..4515,4516..4533,4534..4540)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4548..4564
                     /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     4553..4575
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4572..4588
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4572..4588
                     /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 ccggcagaaa
      181 aaatgactgc tccaaagaag aagcgtaagg taccggcaga aaaaatgact gctccaaaga
      241 agaagcgtaa ggtaccggca gaaaaaatga ctgctccaaa gaagaagcgt aaggtaccgg
      301 tagaaaaaat gagtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat
      361 tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa
      421 catacggaaa acttaccctt aaatttattt gcactactgg aaaactacct gttccatggg
      481 taagtttaaa catatatata ctaactaacc ctgattattt aaattttcag ccaacacttg
      541 tcactacttt ctgttatggt gttcaatgct tctcgagata cccagatcat atgaaacggc
      601 atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaaagaact atatttttca
      661 aagatgacgg gaactacaag acacgtaagt ttaaacagtt cggtactaac taaccataca
      721 tatttaaatt ttcaggtgct gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg
      781 agttaaaagg tattgatttt aaagaagatg gaaacattct tggacacaaa ttggaataca
      841 actataactc acacaatgta tacatcatgg cagacaaaca aaagaatgga atcaaagttg
      901 taagtttaaa catgatttta ctaactaact aatctgattt aaattttcag aacttcaaaa
      961 ttagacacaa cattgaagat ggaagcgttc aactagcaga ccattatcaa caaaatactc
     1021 caattggcga tggccctgtc cttttaccag acaaccatta cctgtccaca caatctgccc
     1081 tttcgaaaga tcccaacgaa aagagagacc acatggtcct tcttgagttt gtaacagctg
     1141 ctgggattac acatggcatg gatgaactat acaaatagca ttcgtagaat tgtaagttta
     1201 aactttctca tactaactaa cgcgctctat ttaaattttc agaattccaa ctgagcgccg
     1261 gtcgctacca ttaccaactt gtctggtgtc aaaaataata ggggccgctg tcatcagagt
     1321 aagtttaaac tgagttctac taactaacga gtaatattta aattttcagc atctcgcgcc
     1381 cgtgcctctg acttctaagt ccaattactc ttcaacatcc ctacatgctc tttctccctg
     1441 tgctcccacc ccctattttt gttattatca aaaaacttct cttaatttct ttgtttttta
     1501 gcttctttta agtcacctct aacaatgaaa ttgtgtagat tcaaaaatag aattaattcg
     1561 taataaaaag tcgaaaaaaa ttgtgctccc tccccccatt aataataatt ctatcccaaa
     1621 atctacacaa tgttctgtgt acacttctta tgttttttac ttctgataaa tttttttgaa
     1681 acatcataga aaaaaccgca cacaaaatac cttatcatat gttacgtttc agtttatgac
     1741 cgcaattttt atttcttcgc acgtctgggc ctctcatgac gtcaaatcat gctcatcgtg
     1801 aaaaagtttt ggagtatttt tggaattttt caatcaagtg aaagtttatg aaattaattt
     1861 tcctgctttt gctttttggg gtttccccta ttgtttgtca agatttcgag gacggcgttt
     1921 ttcttgctaa aatcacaagt attgatgagc acgatgcaag aaagatcgga agaaggtttg
     1981 ggtttgaggc tcagtggaag gtgagtagaa gttgataatt tgaaagtgga gtagtgtcta
     2041 tggggttttt gccttaaatg acagaataca ttcccaatat accaaacata actgtttcct
     2101 actagtcggc cgtacgggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     2161 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     2221 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     2281 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     2341 gtaaggagaa aataccgcat caggcggcct taagggcctc gtgatacgcc tatttttata
     2401 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     2461 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2521 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     2581 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     2641 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     2701 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     2761 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     2821 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     2881 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     2941 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     3001 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3061 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     3121 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     3181 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     3241 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     3301 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     3361 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     3421 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     3481 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3541 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3601 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3661 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3721 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     3781 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3841 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3901 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3961 caccgaactg agatacctac agcgtgagca ttgagaaagc gccacgcttc ccgaagggag
     4021 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4081 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4141 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4201 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     4261 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     4321 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     4381 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     4441 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     4501 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     4561 acaatttcac acaggaaaca gctatgacca tgattacgcc aagctgtaag tttaaacatg
     4621 atcttactaa ctaactattc tcatttaaat tttcagagct taaaaatggc tgaaatcact
     4681 cacaacgatg gatacgctaa caacttggaa atgaaat
//
