LOCUS       sequence-141-exp        8492 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 8492)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1997
REFERENCE   2  (bases 1 to 8492)
  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: L3135
FEATURES             Location/Qualifiers
     source          1..8492
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             527..547
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(590..618)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1366..1386
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1519..1541)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5932..5954)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6054..6073
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6151..6169)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6237..6341
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6342..6410,6411..7202)
                     /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(6560..6579)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7373..7961
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7862..7881
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8115..8132
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8249..8270
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8285..8290,8291..8308,8309..8315)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8323..8339
                     /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     8328..8350
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8347..8363
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8347..8363
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggc cttggtcgac tctagaggat ccccggggat tggccaaagg
       61 acccaaaggt atgtttcgaa tgatactaac ataacataga acattttcag gaggaccctt
      121 gcttggctag caaaaatgca taaggttttg ctggcactgt tctttatctt tctggcacca
      181 gcatccgcac tggcagtctc cgaaccggcc tgcaggatcg attttttgca aattacgagc
      241 gttgtagggg gcggagcgat aggtcctata ggttttggta tatcatcatt cattcattca
      301 ttggtacatt catttaccca ccttcctctt tctgagcttc tctggagttc tgtgcttcct
      361 ttttccctta tctttatact gtaattttta actttcaggc attgattgga tccccgggat
      421 tggccaaagg acccaaaggt atgtttcgaa tgatactaac ataacataga acattttcag
      481 gaggaccctt gcttggaggg taccgagctc agaaaaaatg actgctccaa agaagaagcg
      541 taaggtaccg gtagaaaaaa tgagtaaagg agaagaactt ttcactggag ttgtcccaat
      601 tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga
      661 aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc
      721 tgttccatgg gtaagtttaa acatatatat actaactaac cctgattatt taaattttca
      781 gccaacactt gtcactactt tctgttatgg tgttcaatgc ttctcgagat acccagatca
      841 tatgaaacgg catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac
      901 tatatttttc aaagatgacg ggaactacaa gacacgtaag tttaaacagt tcggtactaa
      961 ctaaccatac atatttaaat tttcaggtgc tgaagtcaag tttgaaggtg atacccttgt
     1021 taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc ttggacacaa
     1081 attggaatac aactataact cacacaatgt atacatcatg gcagacaaac aaaagaatgg
     1141 aatcaaagtt gtaagtttaa acatgatttt actaactaac taatctgatt taaattttca
     1201 gaacttcaaa attagacaca acattgaaga tggaagcgtt caactagcag accattatca
     1261 acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac
     1321 acaatctgcc ctttcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt
     1381 tgtaacagct gctgggatta cacatggcat ggatgaacta tacaaatgcc cggtgggtga
     1441 agaccagaaa cagcacctcg aactgagccg cgatattgcc cagcgtttca acgcgctgta
     1501 tggcgagatc gatcccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     1561 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     1621 caccgatcgc ccttcccaac agttgcgaag gtaagtttaa acagatccat actaactaac
     1681 ttgttctgac ataattttca gcttgaatgg cgaatggcgc tttgcctggt ttccggcacc
     1741 agaagcggtg ccggaaagct ggctggagtg cgatcttcct gaggccgata ctgtcgtcgt
     1801 cccctcaaac tggcagatgc acggttacga tgcgcccatc tacaccaacg tgacctatcc
     1861 cattacggtc aatccgccgt ttgttcccac ggagaatccg acgggttgtt actcgctcac
     1921 atttaatgtt gatgaaagct ggctacagga aggccagacg cgaattattt ttgatggcgt
     1981 taactcggcg tttcatctgt ggtgcaacgg gcgctgggtc ggttacggcc aggtaagttt
     2041 aattaagttg atactaacta acaagatctg attaattttc aggacagtcg tttgccgtct
     2101 gaatttgacc tgagcgcatt tttacgcgcc ggagaaaacc gcctcgcggt gatggtgctg
     2161 cgctggagtg acggcagtta tctggaagat caggatatgt ggcggatgag cggcattttc
     2221 cgtgacgtct cgttgctgca taaaccgact acacaaatca gcgatttcca tgttgccact
     2281 cgctttaatg atgatttctc ccgcgctgta ctggaggctg aagtccaggt aagtttaaac
     2341 aggatcttac taactaacat gctaacactg aattttcaga tgtgcggcga gttgcgtgac
     2401 tacctacggg taacagtttc tttatggcag ggtgaaacgc aggtcgccag cggcaccgcg
     2461 cctttcggcg gtgaaattat cgatgagcgt ggtggttatg ccgatcgcgt cacactacgt
     2521 ctgaacgtcg aaaacccgaa actgtggagc gccgaaatcc cgaatctcta tcgtgcggtg
     2581 gttgaactgc acaccgccga cggcacgctg attgaagcag aagcctgcga tgtcggtttc
     2641 cgcgaggtgc ggattgaaaa tggtctgctg ctgctgaacg gcaagccgtt gctgattcga
     2701 ggcgttaacc gtcacgagca tcatcctctg catggtcagg tcatggatga gcagacgatg
     2761 gtgcaggatg taagtttaaa ctattcgtta ctaactaact ttaaacattt aaattttcag
     2821 atcctgctga tgaagcagaa caactttaac gccgtgcgct gttcgcatta tccgaaccat
     2881 ccgctgtggt acacgctgtg cgaccgctac ggcctgtatg tggtggatga agccaatatt
     2941 gaaacccacg gcatggtgcc aatgaatcgt ctgaccgatg atccgcgctg gctaccggcg
     3001 atgagcgaac gcgtaacgcg aatggtgcag cgcgatcgta atcacccgag tgtgatcatc
     3061 tggtcgctgg ggaatggtaa gtttaaacag ttgaatacta actaacggag atctttgaaa
     3121 ttttcagaat caggccacgg cgctaatcac gacgcgctgt atcgctggat caaatctgtc
     3181 gatccttccc gcccggtgca gtatgaaggc ggcggagccg acaccacggc caccgatatt
     3241 atttgcccga tgtacgcgcg cgtggatgaa gaccagccct tcccggctgt gccgaaatgg
     3301 tccatcaaaa aatggctttc gctacctgga gagacgcgcc cgctgattct ttgcgaggta
     3361 agtttaaaca gaactctact aactaacaca ttagatccta attttcagta cgctcacgcg
     3421 atgggcaaca gtcttggcgg tttcgctaaa tactggcagg cgtttcgtca gtatccccgt
     3481 ttacagggcg gcttcgtctg ggactgggtg gatcagtcgc tgattaaata tgatgaaaac
     3541 ggcaacccgt ggtcggctta cggcggtgat tttggcgata cgccgaacga tcgccagttc
     3601 tgtatgaacg gtctggtctt tgccgaccgc acgccgcatc cagcgtaagt ttaaacaata
     3661 acctactaac taacgtagat aatttaaatt ttcaggctga cggaagcaaa acaccagcag
     3721 cagtttttcc agttccgttt atccgggcaa accatcgaag tgaccagcga atacctgttc
     3781 cgtcatagcg ataacgagct cctgcactgg atggtggcgc tggatggtaa gccgctggca
     3841 agcggtgaag tgcctctgga tgtcgctcca caaggtaaac agttgattga actgcctgaa
     3901 ctaccgcagc cggagagcgc cgggcaactc tggctcacag tacgcgtagt gcaaccgaac
     3961 gcgaccgcat ggtcagaagc cgggcacatc agcgcatggc agcagtggag gtaagtttaa
     4021 acaagatcct actaactaac tctacattga tgaattttca gactggcgga aaacctcagt
     4081 gtgacgctcc ccgccgcgtc ccacgccatc ccgcatctga ccaccagcga aatggatttt
     4141 tgcatcgagc tgggtaataa gcgttggcaa tttaaccgcc agtcaggctt tctttcacag
     4201 atgtggattg gcgataaaaa acaactgctg acgccgctgc gcgatcagtt cacccgtgca
     4261 ccgctggata acgacattgg cgtaagtgaa gcgacccgca ttgaccctaa cgcctgggtc
     4321 ggtaagttta aacaaagttg tactaactaa cgaagatctt gataattttc agaacgctgg
     4381 aaggcggcgg gccattacca ggccgaagca gcgttgttgc agtgcacggc agatacactt
     4441 gctgatgcgg tgctgattac gaccgctcac gcgtggcagc atcaggggaa aaccttattt
     4501 atcagccgga aaacctaccg gattgatggt agtggtcaaa tggcgattac cgttgatgtt
     4561 gaagtggcga gcgatacacc gcatccggcg cggattggcc tgaactgcca gctggcgcag
     4621 gtagcagagc gggtaaactg gctcggatta gggccgcaag aaaactatcc cgaccgcctt
     4681 actgccgcct gttttgaccg ctgggatctg ccattgtcag acatgtagta agtttaaact
     4741 tgatagtact aactaacatg tttcatttaa attttcagta ccccgtacgt cttcccgagc
     4801 gaaaacggtc tgcgctgcgg gacgcgcgaa ttgaattatg gcccacacca gtggcgcggc
     4861 gacttccagt tcaacatcag ccgctacagt caacagcaac tgatggaaac cagccatcgc
     4921 catctgctgc acgcggaaga aggcacatgg ctgaatatcg acggtttcca tatggggatt
     4981 ggtggcgacg actcctggag cccgtcagta tcggcggaat tccaactgag cgccggtcgc
     5041 taccattacc aacttgtctg gtgtcaaaaa taataggggc cgctgtcatc agagtaagtt
     5101 taaactgagt tctactaact aacgagtaat atttaaattt tcagcatctc gcgcccgtgc
     5161 ctctgacttc taagtccaat tactcttcaa catccctaca tgctctttct ccctgtgctc
     5221 ccacccccta tttttgttat tatcaaaaaa cttctcttaa tttctttgtt ttttagcttc
     5281 ttttaagtca cctctaacaa tgaaattgtg tagattcaaa aatagaatta attcgtaata
     5341 aaaagtcgaa aaaaattgtg ctccctcccc ccattaataa taattctatc ccaaaatcta
     5401 cacaatgttc tgtgtacact tcttatgttt tttacttctg ataaattttt ttgaaacatc
     5461 atagaaaaaa ccgcacacaa aataccttat catatgttac gtttcagttt atgaccgcaa
     5521 tttttatttc ttcgcacgtc tgggcctctc atgacgtcaa atcatgctca tcgtgaaaaa
     5581 gttttggagt atttttggaa tttttcaatc aagtgaaagt ttatgaaatt aattttcctg
     5641 cttttgcttt ttggggtttc ccctattgtt tgtcaagatt tcgaggacgg cgtttttctt
     5701 gctaaaatca caagtattga tgagcacgat gcaagaaaga tcggaagaag gtttgggttt
     5761 gaggctcagt ggaaggtgag tagaagttga taatttgaaa gtggagtagt gtctatgggg
     5821 tttttgcctt aaatgacaga atacattccc aatataccaa acataactgt ttcctactag
     5881 tcggccgtac gggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     5941 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     6001 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     6061 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     6121 gagaaaatac cgcatcaggc ggccttaagg gcctcgtgat acgcctattt ttataggtta
     6181 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     6241 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     6301 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     6361 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     6421 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     6481 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     6541 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     6601 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     6661 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     6721 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     6781 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     6841 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     6901 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     6961 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     7021 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     7081 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     7141 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     7201 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     7261 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     7321 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     7381 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     7441 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     7501 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     7561 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     7621 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     7681 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     7741 aactgagata cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg
     7801 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7861 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7921 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7981 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     8041 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     8101 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     8161 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     8221 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     8281 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     8341 ttcacacagg aaacagctat gaccatgatt acgccaagct gtaagtttaa acatgatctt
     8401 actaactaac tattctcatt taaattttca gagcttaaaa atggctgaaa tcactcacaa
     8461 cgatggatac gctaacaact tggaaatgaa at
//
