LOCUS       sequence-290-exp        7515 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 7515)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 7515)
  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: L4796
FEATURES             Location/Qualifiers
     source          1..7515
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3493..3513
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3538..3558
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3601..3629)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4377..4397
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4955..4977)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5077..5096
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5174..5192)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5260..5364
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5365..5433,5434..6225)
                     /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(5583..5602)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6396..6984
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6885..6904
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7138..7155
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7272..7293
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7308..7313,7314..7331,7332..7338)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7346..7362
                     /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     7351..7373
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7370..7386
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7370..7386
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttctgg agattgctcc agcagctctt aacagtgtct acgaccgtct tcgtgcaatt
       61 ctcatggaaa ctgaaagatc ggaaaatgca ctggatcgac gtattcagta tatgattgag
      121 actgcaatgc agattcgaaa ggacaaattt gcggtaaggt agaatatata aatagtttat
      181 tagaaaaaaa taaattagaa taatttaaat tcctactagc caatcaggcg acctttttgc
      241 gcatagttct attattgaaa aatttggaga atttctcata ttctcgctcg gaaatctgga
      301 attcgacgag atcttctggc ttctgtgcag ctgcatcgct ttgtgctccc tttctcgctt
      361 gtcttctgtg tacaccaaga accttgttga gttcatcaac tgaatctgtg actggcttgt
      421 tgctcactgg atgcactaga cgactgattc tcgagaaatc agattgagtt gcgattaggg
      481 tgacctagaa attgggaata atacgaactt ttgaaaatat tcaggaggat taaaaaaatt
      541 attctcgaca atcctacaaa tttacttatt gcaccatgtt gctccaacat ttttcattaa
      601 aagttaatga aaaaatgtag aaaatcggaa attggcaatt ttcagaccat ttttaagcat
      661 tttcaaaaaa aaattgcagc tgaaataaat gtcattttca gataaatcga gcgattttct
      721 gttgtctgac actagttttt agttttaaaa aatgttggaa gaacatggtg caataggtaa
      781 tttcatagaa tttccatgtg ttttttttca attaaccaat tatccaaatc ttccaaactc
      841 acattttgcg gagctgggct atcaagaatc tgctgcagtt ttataagacg agcatctctg
      901 atatcactga aaattaattt ttaatcaaaa cttgaatatc aactaaaccc acttattaac
      961 tttctcgatc ttctgtcgtt cggtacgatg acggtgaaga agccaattgt agtagttgat
     1021 ttggttcaag tcctttcggt gttgtacgtc agtgtcctgc aatgctattt agttataact
     1081 taggcctaag attcaattta atgaagtgat taaatttgtt ctctgaacct cttaagatga
     1141 tcttttggat tagaaacata taagacaggt ttacctatct attaaaaaac agatcaaaat
     1201 agatacgacc aaatcggata atccatgcct acctggcatc taggaacgtg ttcttagaag
     1261 atttcttacg taatcgtatg aagaaataac aatttgatcg ttggccagca aaaatagggt
     1321 tttaagtggg atagtgtttt tattagctaa ccggaaaatt ttatagtttt tttttgcaag
     1381 aaaccactga aaacccccta attgtataca ttttttggag cagcttctgg tctttttgag
     1441 caataaaatt cgataaaaca gaatttaagt gtaaattgtt cacatttagt ttctatttta
     1501 tcaaattttg ttgctcaaaa acattcgaag ctgctctaaa aaaatgcatt aaaaaagggg
     1561 ttttcagtgg tttttcacat taaaaaagct aattttaact aaaaatccat catatttcca
     1621 actttgtcac aacaataaaa tgctggtcaa aatgtgttcg aaaaaatgtt ttttttttta
     1681 atttttataa tttaaaaata gttttctttc gctgggacac atacattttt gggcgtaaat
     1741 tttcagttca aatttccatt tttacaacca taatcataaa gctacgtctg atctctctcg
     1801 cacttacctg cgcctgattc gaaagaacaa ccgtagccaa aagaacaaga agaacaagca
     1861 cgtagttgtg gtagtggacg ttcatcacgc aatactgacc aatggtcgtg gggtctcact
     1921 ttccgtacta ttgagagagg ggagactgaa gatggcaatt gaggacagtg tcttcgacgc
     1981 acgcatgcat ccataagcat aatccaggag ggatggagag aaaaatcttg tttctaagcc
     2041 cctccctttg taatacatac acatatctaa taccgaagaa tggctaattg aatggacgtc
     2101 agctgttgct gtagttgcca aggcatcatc gatgaaataa ctgaaagaaa gaattaaata
     2161 attattgcag gcgtatccgg cggtcattga agacttggac ttgattgagg aggaggatca
     2221 gatcatccat acacttaatt tggaggatgc ggttgatccg gaaaatgggc ttagtaagtg
     2281 actgaccaca cgcggggggc attaatttaa taaattgaat tccatttcag atgtgttcaa
     2341 actagatcca gaattcgaaa agaacgagga ggtttatgag gagatccgta aggaaatcat
     2401 tggaaacgcc gatatttcgg atgaggatgg tggcgacgag ttggatgatg aagaagaggg
     2461 tagtgatgtg gaagaggctc cgaagaagac tacagagatt attgataata ctgatcagaa
     2521 ttgactgctt tcagaaggta ttcattttga gttttgggcc ggcaaatctg taagttgccg
     2581 gttgccgaaa atttgctgaa tttgccggaa aaaaaaattc cggaatttat ttaaaaactt
     2641 tttgtaaaaa ttaaattaaa tttgcaactt ttcagagaag tctacctgac aatgcaatca
     2701 tctttggact accaagaagc tgctcacaaa ttgctgaaaa tgaagattcc agacagcatg
     2761 caggtcagcg atgttgcaaa gaaaaatttt cgaccaaaaa aaccaaccaa tcataaaatt
     2821 taaaaaaaaa ctccgttttt ttcttttttt ttatacgaga aaaaccaaaa aaatgtattt
     2881 ttgccaaatt ctaaaatact atccccgaaa ttttcaatat tttctctttc agaacgaact
     2941 ctgcgcgatg cttgtcgatt gttgtgctca acagcgtacc tacgagcgat tctacggaat
     3001 gctgagaaca cgttaaacac gaagatcata tttattttga gacccggatg ctctgaaaat
     3061 gtctgacata gatttaaaaa agcatatata tatttttcat tttcaacgtg aaagttttgt
     3121 gcaactttat agaatctcct attggcacat tgttttttat ttaactgagg cagtttttga
     3181 acaccttttt gaaactttga atctctttga agtatactgt cgaaaagact gacttgagcg
     3241 ttcgaaatgc cagaagaaaa ctatatttga atctcgcgct aaattgagaa atgcaaccgc
     3301 gctccactgg acaattggaa aaaaaattta ttcggaggcg acaacggtat tttcgaaatt
     3361 gattttctgt gtattttctc attttttata aattcttctt tgatttatcg ttcgtttgtg
     3421 agaaatttaa ttgtattcaa acttttttat agtaagatac cggtggtacc gagctcagaa
     3481 aaaatgaccg ctccaaagaa gaaacgcaaa gtaccgagct cagaaaaaat gacggcacct
     3541 aaaaagaaac gcaaagtacc ggtagaaaaa atgagtaaag gagaagaact tttcactgga
     3601 gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtcagt
     3661 ggagagggtg aaggtgatgc aacatacgga aaacttaccc ttaaatttat ttgcactact
     3721 ggaaaactac ctgttccatg ggtaagttta aacatatata tactaactaa ccctgattat
     3781 ttaaattttc agccaacact tgtcactact ttctgttatg gtgttcaatg cttctcgaga
     3841 tacccagatc atatgaaacg gcatgacttt ttcaagagtg ccatgcccga aggttatgta
     3901 caggaaagaa ctatattttt caaagatgac gggaactaca agacacgtaa gtttaaacag
     3961 ttcggtacta actaaccata catatttaaa ttttcaggtg ctgaagtcaa gtttgaaggt
     4021 gatacccttg ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt
     4081 cttggacaca aattggaata caactataac tcacacaatg tatacatcat ggcagacaaa
     4141 caaaagaatg gaatcaaagt tgtaagttta aacatgattt tactaactaa ctaatctgat
     4201 ttaaattttc agaacttcaa aattagacac aacattgaag atggaagcgt tcaactagca
     4261 gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc agacaaccat
     4321 tacctgtcca cacaatctgc cctttcgaaa gatcccaacg aaaagagaga ccacatggtc
     4381 cttcttgagt ttgtaacagc tgctgggatt acacatggca tggacgaact atacaaaagc
     4441 ccacaagctt tggaattcgc tagccggcca tacaagtaat ccggatgatc gacgccaacg
     4501 tcgttgaatt ttcaaatttt aaatactgaa tatttgtttt ttttcctatt atttatttat
     4561 tctctttgtg ttttttttct tgctttctaa aaaattaatt caatccaaat ctaaacattt
     4621 ttttttctct ttccgtctcc caattcgtat tccgctcctc tcatctgaac acaatgtgca
     4681 agtttattta tcttctcgct ttcatttcat taggacgtgg ggggaattgg tggaaggggg
     4741 aaacacacaa aaggatgatg gaaatgaaat aaggacacac aatatgcaac aacattcaat
     4801 tcagaaatat ggaggaaggt ttaaaagaaa acataaaaat atatagagga ggaaggaaaa
     4861 ctagtaaaaa ataagcaaag aaattaggcg aacgatgaga attgtcctcg cttgggccct
     4921 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     4981 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     5041 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     5101 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     5161 ggcggcctta agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     5221 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5281 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5341 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5401 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5461 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5521 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5581 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5641 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5701 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5761 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5821 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5881 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5941 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6001 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6061 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6121 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6181 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6241 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6301 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6361 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6421 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6481 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6541 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6601 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6661 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6721 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6781 cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6841 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6901 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6961 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7021 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     7081 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     7141 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     7201 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     7261 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     7321 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     7381 tatgaccatg attacgccaa gctgtaagtt taaacatgat cttactaact aactattctc
     7441 atttaaattt tcagagctta aaaatggctg aaatcactca caacgatgga tacgctaaca
     7501 acttggaaat gaaat
//
