LOCUS       sequence-263-exp        6591 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 6591)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 6591)
  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: L4438
FEATURES             Location/Qualifiers
     source          1..6591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             503..523
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4165..4187)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4287..4306
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4384..4402)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4470..4574
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4575..4643,4644..5435)
                     /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(4793..4812)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5606..6194
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6095..6114
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6348..6365
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6482..6503
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6518..6523,6524..6541,6542..6548)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6556..6572
                     /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     6561..6583
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6580..6591,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6580..6591,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccgggta
       61 cctaatttga tcgataatat gtacatattt tgttattcgt tccggtatcg tacctccacc
      121 taccataaat cccataccct acatctttat cgattttttg caaattacga gcgttgtagg
      181 gggcggagcg ataggtccta taggttttgg tatatcatca ttcattcatt cattggtaca
      241 ttcatttacc caccttcctc tttctgagct tctctggagt tctgtgcttc ctttttccct
      301 tatctttata ctgtaatttt taactttcag gcattgattg gatccccggg attggccaaa
      361 ggacccaaag gtatgtttcg aatgatacta acataacata gaacattttc aggaggaccc
      421 ttggctagag gatccccggg attggccaaa ggacccaaag gaggaccctt ggagggtacc
      481 gagctcagaa aaaatgactg ctccaaagaa gaagcgtaag gtaccggtag aaaaaatgaa
      541 catcatcgaa aacatcgaaa agaatgactt ctcagaaatc gaactggctg ctatcccgtt
      601 caacacactg gctgaccact acggaagcgc cttggctaaa gagcagttgg ctttagaaca
      661 tgagtcttat gagctaggcg agcgccgctt cctcaagatg cttgagcgtc aagcgaaagc
      721 tggtgagatt gcagacaacg cagccgctaa gccgttactc gctacgcttc tccctaagtt
      781 aaccacacgt atcgtcgagt ggctcgaaga gtacgcatcg aagaaaggcc gcaagcctag
      841 cgcatacgca ccgctccagt tactcaagcc ggaggcctcc gcgtttatca ccctgaaagt
      901 tatccttgcg tcactaacca gtacgaacat gacaaccatt caggccgctg ctggtatgct
      961 ggggaaagcc attgaggacg aggcacgatt tgggcgcatc cgtgacctag aagcgaagca
     1021 cttcaagaag cacgttgagg aacagcttaa caagcgccac gggcaagtct acaagaaagc
     1081 atttatgcag gtggtcgagg ccgatatgat tggtcgaggt ctgcttggtg gcgaggcgtg
     1141 gtctagctgg gataaagaaa ccacgatgca cgtagggatt cgcctgattg aaatgctgat
     1201 tgaatccacg ggtctggtgg aattacagcg ccacaacgca ggtaacgcag gctctgacca
     1261 tgaggcactg caactggccc aagagtacgt ggacgtatta gcgaagcgtg caggcgctct
     1321 ggcgggtatc tctccgatgt tccagccgtg tgtcgtaccg ccgaaacctt gggtagcaat
     1381 cacagggggc ggctattggg ctaacggtcg cagacctttg gcactcgttc gcactcactc
     1441 taagaagggc ttgatgcgct acgaagacgt ttacatgcca gaagtctaca aggctgtgaa
     1501 cctcgcgcaa aacaccgcat ggaaaatcaa caagaaagtt cttgctgttg tcaatgagat
     1561 tgttaactgg aagaattgcc cggtagcaga cattccatcg ctggagcgcc aagagttacc
     1621 gcctaagcct gacgacattg acaccaacga ggcagcgctc aaggagtgga agaaagccgc
     1681 tgctggtatc tatcgcttgg acaaggcacg agtgtctcgc cgtatcagct tagagttcat
     1741 gctggagcag gccaacaagt tcgcaagcaa gaaaggtaag tttaaacttg gatctactaa
     1801 ctaacgaatc catttaaatt tttcagcaat ctggttccct tacaacatgg actggcgcgg
     1861 tcgtgtgtac gctgtgccga tgttcaaccc gcaaggcaac gacatgacga aaggtctgct
     1921 gacccttgct aaaggcaagc caatcggtga ggaaggtttc tactggctga aaatccacgg
     1981 tgcgaactgt gcgggtgttg ataaggttcc attcccggag cgcatcgcgt tcattgagaa
     2041 gcacgtagac gacattctgg cttgcgctaa agacccaatc aataacactt ggtgggctga
     2101 gcaggattca ccgttctgtt tcctcgcgtt ttgcttcgag tatgcaggcg ttacgcacca
     2161 cggtctgagc tacaattgct ctctgccgct ggcgttcgac gggtcttgct ctggtatcca
     2221 gcacttctcc gcgatgctcc gcgatgaggt aggcggtcgt gcggttaacc tgctgccaag
     2281 cgaaaccgtg caggacattt acggcatcgt tgcacagaaa gtaaacgaga ttctcaaaca
     2341 ggatgcaatc aacggcacgc ctaacgagat gattaccgtg accgacaagg acaccgggga
     2401 aatctcagag aagctcaaac ttggaacctc aacgctggcg caacagtggc tggcatatgg
     2461 tgtaacccgt agcgtaacta aacgttcggt catgacgctg gcttacggtt ccaaggagtt
     2521 cggctttcgt caacaggtat tggatgacac cattcagcct gcaattgaca gcggtaaggg
     2581 cttgatgttc acccaaccga accaagcggc tggctatatg gctaagctga tttgggatgc
     2641 ggtaagcgtg accgtagttg cagcggttga ggcgatgaac tggctcaaat ctgccgctaa
     2701 gctgctggct gctgaggtca aggacaagaa gaccaaggag attctgcgcc accgttgcgc
     2761 ggttcactgg actacgccgg acggcttccc ggtctggcag gaataccgca agccactcca
     2821 gaagcgtctc gatatgattt tcttagggca attccgtctg caaccgacga ttaataccct
     2881 caaggattca ggcattgacg cacacaagca ggagtctggc atcgctccta actttgttca
     2941 ctcacaggac ggtagccacc tccgcatgac agtcgtttat gctcacgaga agtatggcat
     3001 tgagtccttt gcgctcatcc atgacagctt tgggactatc ccggcagacg ctggtaagct
     3061 ctttaaggct gtgcgtgaaa cgatggttat cacctatgag aacaacgatg tgctggcaga
     3121 cttctactct cagtttgccg accagctaca cgagacccaa ctggacaaga tgcctccgct
     3181 tccgaagaaa ggaaacctga acctgcaaga cattctcaag tctgactttg cctttgcata
     3241 gcattcgtag aattccaact gagcgccggt cgctaccatt accaacttgt ctggtgtcaa
     3301 aaataatagg ggccgctgtc atcagagtaa gtttaaactg agttctacta actaacgagt
     3361 aatatttaaa ttttcagcat ctcgcgcccg tgcctctgac ttctaagtcc aattactctt
     3421 caacatccct acatgctctt tctccctgtg ctcccacccc ctatttttgt tattatcaaa
     3481 aaacttctct taatttcttt gttttttagc ttcttttaag tcacctctaa caatgaaatt
     3541 gtgtagattc aaaaatagaa ttaattcgta ataaaaagtc gaaaaaaatt gtgctccctc
     3601 cccccattaa taataattct atcccaaaat ctacacaatg ttctgtgtac acttcttatg
     3661 ttttttactt ctgataaatt tttttgaaac atcatagaaa aaaccgcaca caaaatacct
     3721 tatcatatgt tacgtttcag tttatgaccg caatttttat ttcttcgcac gtctgggcct
     3781 ctcatgacgt caaatcatgc tcatcgtgaa aaagttttgg agtatttttg gaatttttca
     3841 atcaagtgaa agtttatgaa attaattttc ctgcttttgc tttttggggt ttcccctatt
     3901 gtttgtcaag atttcgagga cggcgttttt cttgctaaaa tcacaagtat tgatgagcac
     3961 gatgcaagaa agatcggaag aaggtttggg tttgaggctc agtggaaggt gagtagaagt
     4021 tgataatttg aaagtggagt agtgtctatg gggtttttgc cttaaatgac agaatacatt
     4081 cccaatatac caaacataac tgtttcctac tagtcggccg tacgggccct ttcgtctcgc
     4141 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     4201 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     4261 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     4321 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcggcctta
     4381 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     4441 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4501 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4561 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4621 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4681 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4741 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4801 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4861 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     4921 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4981 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5041 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5101 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5161 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5221 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5281 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5341 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5401 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5461 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5521 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5581 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5641 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5701 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     5761 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5821 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5881 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5941 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagcatt
     6001 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6061 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6121 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6181 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6241 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6301 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     6361 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     6421 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     6481 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     6541 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc t
//
