LOCUS       sequence-127-exp        8407 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 8407)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1997
REFERENCE   2  (bases 1 to 8407)
  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: L2681
FEATURES             Location/Qualifiers
     source          1..8407
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             576..596
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(639..667)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1415..1435
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1568..1590)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5981..6003)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6103..6122
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6200..6218)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6286..6390
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6391..6459,6460..7251)
                     /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(6609..6628)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7422..8010
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7911..7930
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8164..8181
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8298..8319
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8334..8339,8340..8357,8358..8364)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8372..8388
                     /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     8377..8399
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(8396..8407,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(8396..8407,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tcaagagcat
       61 ttgaatcaga atatggagaa cggagcatga gcattttcga agttttttag atgcactaga
      121 acaaagcgtg ttggcttcct ctgagcccgc tttccttata tacccgcatt ctgcagcctt
      181 acagaatgtt ctagaaggtc ctagatgcat tcgtttgaaa atactcccgg tgggtgcaaa
      241 gagacgcaga cggaaaatgt atctgggtct ctttattgtg tacactactt ttccatgtac
      301 cgaatgtgag tcgccctcct tttgcaacaa gcagctcgaa tgttctagaa aaaggtggaa
      361 aatagtataa ataccgttga aaataaatac cgaacaacat ttgctctaat tgtgaaatta
      421 gaaatcttca aactataatc atctcactgg atccccggga ttggccaaag gacccaaagg
      481 tatgtttcga atgatactaa cataacatag aacattttca ggaggaccct tggctagagg
      541 atccccgggt accgagctca gaaaaaatga ctgctccaaa gaagaagcgt aaggtaccgg
      601 tagaaaaaat gagtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat
      661 tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa
      721 catacggaaa acttaccctt aaatttattt gcactactgg aaaactacct gttccatggg
      781 taagtttaaa catatatata ctaactaacc ctgattattt aaattttcag ccaacacttg
      841 tcactacttt ctgttatggt gttcaatgct tctcgagata cccagatcat atgaaacggc
      901 atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaaagaact atatttttca
      961 aagatgacgg gaactacaag acacgtaagt ttaaacagtt cggtactaac taaccataca
     1021 tatttaaatt ttcaggtgct gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg
     1081 agttaaaagg tattgatttt aaagaagatg gaaacattct tggacacaaa ttggaataca
     1141 actataactc acacaatgta tacatcatgg cagacaaaca aaagaatgga atcaaagttg
     1201 taagtttaaa catgatttta ctaactaact aatctgattt aaattttcag aacttcaaaa
     1261 ttagacacaa cattgaagat ggaagcgttc aactagcaga ccattatcaa caaaatactc
     1321 caattggcga tggccctgtc cttttaccag acaaccatta cctgtccaca caatctgccc
     1381 tttcgaaaga tcccaacgaa aagagagacc acatggtcct tcttgagttt gtaacagctg
     1441 ctgggattac acatggcatg gatgaactat acaaatgccc ggtgggtgaa gaccagaaac
     1501 agcacctcga actgagccgc gatattgccc agcgtttcaa cgcgctgtat ggcgagatcg
     1561 atcccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     1621 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     1681 cttcccaaca gttgcgaagg taagtttaaa cagatccata ctaactaact tgttctgaca
     1741 taattttcag cttgaatggc gaatggcgct ttgcctggtt tccggcacca gaagcggtgc
     1801 cggaaagctg gctggagtgc gatcttcctg aggccgatac tgtcgtcgtc ccctcaaact
     1861 ggcagatgca cggttacgat gcgcccatct acaccaacgt gacctatccc attacggtca
     1921 atccgccgtt tgttcccacg gagaatccga cgggttgtta ctcgctcaca tttaatgttg
     1981 atgaaagctg gctacaggaa ggccagacgc gaattatttt tgatggcgtt aactcggcgt
     2041 ttcatctgtg gtgcaacggg cgctgggtcg gttacggcca ggtaagttta attaagttga
     2101 tactaactaa caagatctga ttaattttca ggacagtcgt ttgccgtctg aatttgacct
     2161 gagcgcattt ttacgcgccg gagaaaaccg cctcgcggtg atggtgctgc gctggagtga
     2221 cggcagttat ctggaagatc aggatatgtg gcggatgagc ggcattttcc gtgacgtctc
     2281 gttgctgcat aaaccgacta cacaaatcag cgatttccat gttgccactc gctttaatga
     2341 tgatttctcc cgcgctgtac tggaggctga agtccaggta agtttaaaca ggatcttact
     2401 aactaacatg ctaacactga attttcagat gtgcggcgag ttgcgtgact acctacgggt
     2461 aacagtttct ttatggcagg gtgaaacgca ggtcgccagc ggcaccgcgc ctttcggcgg
     2521 tgaaattatc gatgagcgtg gtggttatgc cgatcgcgtc acactacgtc tgaacgtcga
     2581 aaacccgaaa ctgtggagcg ccgaaatccc gaatctctat cgtgcggtgg ttgaactgca
     2641 caccgccgac ggcacgctga ttgaagcaga agcctgcgat gtcggtttcc gcgaggtgcg
     2701 gattgaaaat ggtctgctgc tgctgaacgg caagccgttg ctgattcgag gcgttaaccg
     2761 tcacgagcat catcctctgc atggtcaggt catggatgag cagacgatgg tgcaggatgt
     2821 aagtttaaac tattcgttac taactaactt taaacattta aattttcaga tcctgctgat
     2881 gaagcagaac aactttaacg ccgtgcgctg ttcgcattat ccgaaccatc cgctgtggta
     2941 cacgctgtgc gaccgctacg gcctgtatgt ggtggatgaa gccaatattg aaacccacgg
     3001 catggtgcca atgaatcgtc tgaccgatga tccgcgctgg ctaccggcga tgagcgaacg
     3061 cgtaacgcga atggtgcagc gcgatcgtaa tcacccgagt gtgatcatct ggtcgctggg
     3121 gaatggtaag tttaaacagt tgaatactaa ctaacggaga tctttgaaat tttcagaatc
     3181 aggccacggc gctaatcacg acgcgctgta tcgctggatc aaatctgtcg atccttcccg
     3241 cccggtgcag tatgaaggcg gcggagccga caccacggcc accgatatta tttgcccgat
     3301 gtacgcgcgc gtggatgaag accagccctt cccggctgtg ccgaaatggt ccatcaaaaa
     3361 atggctttcg ctacctggag agacgcgccc gctgattctt tgcgaggtaa gtttaaacag
     3421 aactctacta actaacacat tagatcctaa ttttcagtac gctcacgcga tgggcaacag
     3481 tcttggcggt ttcgctaaat actggcaggc gtttcgtcag tatccccgtt tacagggcgg
     3541 cttcgtctgg gactgggtgg atcagtcgct gattaaatat gatgaaaacg gcaacccgtg
     3601 gtcggcttac ggcggtgatt ttggcgatac gccgaacgat cgccagttct gtatgaacgg
     3661 tctggtcttt gccgaccgca cgccgcatcc agcgtaagtt taaacaataa cctactaact
     3721 aacgtagata atttaaattt tcaggctgac ggaagcaaaa caccagcagc agtttttcca
     3781 gttccgttta tccgggcaaa ccatcgaagt gaccagcgaa tacctgttcc gtcatagcga
     3841 taacgagctc ctgcactgga tggtggcgct ggatggtaag ccgctggcaa gcggtgaagt
     3901 gcctctggat gtcgctccac aaggtaaaca gttgattgaa ctgcctgaac taccgcagcc
     3961 ggagagcgcc gggcaactct ggctcacagt acgcgtagtg caaccgaacg cgaccgcatg
     4021 gtcagaagcc gggcacatca gcgcatggca gcagtggagg taagtttaaa caagatccta
     4081 ctaactaact ctacattgat gaattttcag actggcggaa aacctcagtg tgacgctccc
     4141 cgccgcgtcc cacgccatcc cgcatctgac caccagcgaa atggattttt gcatcgagct
     4201 gggtaataag cgttggcaat ttaaccgcca gtcaggcttt ctttcacaga tgtggattgg
     4261 cgataaaaaa caactgctga cgccgctgcg cgatcagttc acccgtgcac cgctggataa
     4321 cgacattggc gtaagtgaag cgacccgcat tgaccctaac gcctgggtcg gtaagtttaa
     4381 acaaagttgt actaactaac gaagatcttg ataattttca gaacgctgga aggcggcggg
     4441 ccattaccag gccgaagcag cgttgttgca gtgcacggca gatacacttg ctgatgcggt
     4501 gctgattacg accgctcacg cgtggcagca tcaggggaaa accttattta tcagccggaa
     4561 aacctaccgg attgatggta gtggtcaaat ggcgattacc gttgatgttg aagtggcgag
     4621 cgatacaccg catccggcgc ggattggcct gaactgccag ctggcgcagg tagcagagcg
     4681 ggtaaactgg ctcggattag ggccgcaaga aaactatccc gaccgcctta ctgccgcctg
     4741 ttttgaccgc tgggatctgc cattgtcaga catgtagtaa gtttaaactt gatagtacta
     4801 actaacatgt ttcatttaaa ttttcagtac cccgtacgtc ttcccgagcg aaaacggtct
     4861 gcgctgcggg acgcgcgaat tgaattatgg cccacaccag tggcgcggcg acttccagtt
     4921 caacatcagc cgctacagtc aacagcaact gatggaaacc agccatcgcc atctgctgca
     4981 cgcggaagaa ggcacatggc tgaatatcga cggtttccat atggggattg gtggcgacga
     5041 ctcctggagc ccgtcagtat cggcggaatt ccaactgagc gccggtcgct accattacca
     5101 acttgtctgg tgtcaaaaat aataggggcc gctgtcatca gagtaagttt aaactgagtt
     5161 ctactaacta acgagtaata tttaaatttt cagcatctcg cgcccgtgcc tctgacttct
     5221 aagtccaatt actcttcaac atccctacat gctctttctc cctgtgctcc caccccctat
     5281 ttttgttatt atcaaaaaac ttctcttaat ttctttgttt tttagcttct tttaagtcac
     5341 ctctaacaat gaaattgtgt agattcaaaa atagaattaa ttcgtaataa aaagtcgaaa
     5401 aaaattgtgc tccctccccc cattaataat aattctatcc caaaatctac acaatgttct
     5461 gtgtacactt cttatgtttt ttacttctga taaatttttt tgaaacatca tagaaaaaac
     5521 cgcacacaaa ataccttatc atatgttacg tttcagttta tgaccgcaat ttttatttct
     5581 tcgcacgtct gggcctctca tgacgtcaaa tcatgctcat cgtgaaaaag ttttggagta
     5641 tttttggaat ttttcaatca agtgaaagtt tatgaaatta attttcctgc ttttgctttt
     5701 tggggtttcc cctattgttt gtcaagattt cgaggacggc gtttttcttg ctaaaatcac
     5761 aagtattgat gagcacgatg caagaaagat cggaagaagg tttgggtttg aggctcagtg
     5821 gaaggtgagt agaagttgat aatttgaaag tggagtagtg tctatggggt ttttgcctta
     5881 aatgacagaa tacattccca atataccaaa cataactgtt tcctactagt cggccgtacg
     5941 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     6001 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     6061 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     6121 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     6181 gcatcaggcg gccttaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     6241 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6301 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     6361 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     6421 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     6481 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     6541 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     6601 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     6661 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6721 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6781 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     6841 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6901 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6961 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     7021 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     7081 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     7141 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     7201 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     7261 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     7321 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     7381 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     7441 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     7501 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     7561 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     7621 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7681 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7741 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7801 ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7861 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7921 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7981 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     8041 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     8101 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     8161 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     8221 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     8281 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     8341 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     8401 aacagct
//
