LOCUS       sequence-191-exp        8217 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 8217)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1997
REFERENCE   2  (bases 1 to 8217)
  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: L3617
FEATURES             Location/Qualifiers
     source          1..8217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             386..406
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(449..477)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1225..1245
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1378..1400)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5791..5813)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5913..5932
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6010..6028)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6096..6200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6201..6269,6270..7061)
                     /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(6419..6438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7232..7820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7721..7740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7974..7991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8108..8129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8144..8149,8150..8167,8168..8174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8182..8198
                     /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     8187..8209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(8206..8217,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(8206..8217,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccaattc
       61 ttgagacttc cactatcttc tctggcaccc tccactacgc gtaggcgtct ctcgcttcgt
      121 gtattcccgg gaagccgggt tcccgtctct cccgccgctg ccgctgccgc acacagcttt
      181 acacctcgta gaatccccaa agaggggcgt ggcttgcggg tgccaacatc ctcctgccga
      241 ggaagaagca ggcactcatc actcgcatca tcaacctcgg gattggccaa aggacccaaa
      301 ggtatgtttc gaatgatact aacataacat agaacatttt caggaggacc cttggagggt
      361 accgagctca gaaaaaatga ctgctccaaa gaagaagcgt aaggtaccgg tagaaaaaat
      421 gagtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga
      481 tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa
      541 acttaccctt aaatttattt gcactactgg aaaactacct gttccatggg taagtttaaa
      601 catatatata ctaactaacc ctgattattt aaattttcag ccaacacttg tcactactct
      661 cacttatggt gttcaatgct tctcgagata cccagatcat atgaaacagc atgacttttt
      721 caagagtgcc atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg
      781 gaactacaag acacgtaagt ttaaacagtt cggtactaac taaccataca tatttaaatt
      841 ttcaggtgct gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg agttaaaagg
      901 tattgatttt aaagaagatg gaaacattct tggacacaaa ttggaataca acttcaactc
      961 acacaacgta tacatcactg ccgacaaaca aaagaatggg atcaaagcgg taagtttaaa
     1021 catgatttta ctaactaact aatctgattt aaattttcag aacttcaaaa ttagacacaa
     1081 cattgaagat ggaagcgttc aactagcaga ccattatcaa caaaatactc caattggcga
     1141 tggccctgtc cttttaccag acaaccatta cctgtccaca caatctgccc tttcgaaaga
     1201 tcccaacgaa aagagagacc acatggtcct tcttgagttt gtaacagctg ctgggattac
     1261 acatggcatg gatgaactat acaaatgccc ggtgggtgaa gaccagaaac agcacctcga
     1321 actgagccgc gatattgccc agcgtttcaa cgcgctgtat ggcgagatcg atcccgtcgt
     1381 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     1441 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     1501 gttgcgaagg taagtttaaa cagatccata ctaactaact tgttctgaca taattttcag
     1561 cttgaatggc gaatggcgct ttgcctggtt tccggcacca gaagcggtgc cggaaagctg
     1621 gctggagtgc gatcttcctg aggccgatac tgtcgtcgtc ccctcaaact ggcagatgca
     1681 cggttacgat gcgcccatct acaccaacgt gacctatccc attacggtca atccgccgtt
     1741 tgttcccacg gagaatccga cgggttgtta ctcgctcaca tttaatgttg atgaaagctg
     1801 gctacaggaa ggccagacgc gaattatttt tgatggcgtt aactcggcgt ttcatctgtg
     1861 gtgcaacggg cgctgggtcg gttacggcca ggtaagttta attaagttga tactaactaa
     1921 caagatctga ttaattttca ggacagtcgt ttgccgtctg aatttgacct gagcgcattt
     1981 ttacgcgccg gagaaaaccg cctcgcggtg atggtgctgc gctggagtga cggcagttat
     2041 ctggaagatc aggatatgtg gcggatgagc ggcattttcc gtgacgtctc gttgctgcat
     2101 aaaccgacta cacaaatcag cgatttccat gttgccactc gctttaatga tgatttctcc
     2161 cgcgctgtac tggaggctga agtccaggta agtttaaaca ggatcttact aactaacatg
     2221 ctaacactga attttcagat gtgcggcgag ttgcgtgact acctacgggt aacagtttct
     2281 ttatggcagg gtgaaacgca ggtcgccagc ggcaccgcgc ctttcggcgg tgaaattatc
     2341 gatgagcgtg gtggttatgc cgatcgcgtc acactacgtc tgaacgtcga aaacccgaaa
     2401 ctgtggagcg ccgaaatccc gaatctctat cgtgcggtgg ttgaactgca caccgccgac
     2461 ggcacgctga ttgaagcaga agcctgcgat gtcggtttcc gcgaggtgcg gattgaaaat
     2521 ggtctgctgc tgctgaacgg caagccgttg ctgattcgag gcgttaaccg tcacgagcat
     2581 catcctctgc atggtcaggt catggatgag cagacgatgg tgcaggatgt aagtttaaac
     2641 tattcgttac taactaactt taaacattta aattttcaga tcctgctgat gaagcagaac
     2701 aactttaacg ccgtgcgctg ttcgcattat ccgaaccatc cgctgtggta cacgctgtgc
     2761 gaccgctacg gcctgtatgt ggtggatgaa gccaatattg aaacccacgg catggtgcca
     2821 atgaatcgtc tgaccgatga tccgcgctgg ctaccggcga tgagcgaacg cgtaacgcga
     2881 atggtgcagc gcgatcgtaa tcacccgagt gtgatcatct ggtcgctggg gaatggtaag
     2941 tttaaacagt tgaatactaa ctaacggaga tctttgaaat tttcagaatc aggccacggc
     3001 gctaatcacg acgcgctgta tcgctggatc aaatctgtcg atccttcccg cccggtgcag
     3061 tatgaaggcg gcggagccga caccacggcc accgatatta tttgcccgat gtacgcgcgc
     3121 gtggatgaag accagccctt cccggctgtg ccgaaatggt ccatcaaaaa atggctttcg
     3181 ctacctggag agacgcgccc gctgattctt tgcgaggtaa gtttaaacag aactctacta
     3241 actaacacat tagatcctaa ttttcagtac gctcacgcga tgggcaacag tcttggcggt
     3301 ttcgctaaat actggcaggc gtttcgtcag tatccccgtt tacagggcgg cttcgtctgg
     3361 gactgggtgg atcagtcgct gattaaatat gatgaaaacg gcaacccgtg gtcggcttac
     3421 ggcggtgatt ttggcgatac gccgaacgat cgccagttct gtatgaacgg tctggtcttt
     3481 gccgaccgca cgccgcatcc agcgtaagtt taaacaataa cctactaact aacgtagata
     3541 atttaaattt tcaggctgac ggaagcaaaa caccagcagc agtttttcca gttccgttta
     3601 tccgggcaaa ccatcgaagt gaccagcgaa tacctgttcc gtcatagcga taacgagctc
     3661 ctgcactgga tggtggcgct ggatggtaag ccgctggcaa gcggtgaagt gcctctggat
     3721 gtcgctccac aaggtaaaca gttgattgaa ctgcctgaac taccgcagcc ggagagcgcc
     3781 gggcaactct ggctcacagt acgcgtagtg caaccgaacg cgaccgcatg gtcagaagcc
     3841 gggcacatca gcgcatggca gcagtggagg taagtttaaa caagatccta ctaactaact
     3901 ctacattgat gaattttcag actggcggaa aacctcagtg tgacgctccc cgccgcgtcc
     3961 cacgccatcc cgcatctgac caccagcgaa atggattttt gcatcgagct gggtaataag
     4021 cgttggcaat ttaaccgcca gtcaggcttt ctttcacaga tgtggattgg cgataaaaaa
     4081 caactgctga cgccgctgcg cgatcagttc acccgtgcac cgctggataa cgacattggc
     4141 gtaagtgaag cgacccgcat tgaccctaac gcctgggtcg gtaagtttaa acaaagttgt
     4201 actaactaac gaagatcttg ataattttca gaacgctgga aggcggcggg ccattaccag
     4261 gccgaagcag cgttgttgca gtgcacggca gatacacttg ctgatgcggt gctgattacg
     4321 accgctcacg cgtggcagca tcaggggaaa accttattta tcagccggaa aacctaccgg
     4381 attgatggta gtggtcaaat ggcgattacc gttgatgttg aagtggcgag cgatacaccg
     4441 catccggcgc ggattggcct gaactgccag ctggcgcagg tagcagagcg ggtaaactgg
     4501 ctcggattag ggccgcaaga aaactatccc gaccgcctta ctgccgcctg ttttgaccgc
     4561 tgggatctgc cattgtcaga catgtagtaa gtttaaactt gatagtacta actaacatgt
     4621 ttcatttaaa ttttcagtac cccgtacgtc ttcccgagcg aaaacggtct gcgctgcggg
     4681 acgcgcgaat tgaattatgg cccacaccag tggcgcggcg acttccagtt caacatcagc
     4741 cgctacagtc aacagcaact gatggaaacc agccatcgcc atctgctgca cgcggaagaa
     4801 ggcacatggc tgaatatcga cggtttccat atggggattg gtggcgacga ctcctggagc
     4861 ccgtcagtat cggcggaatt ccaactgagc gccggtcgct accattacca acttgtctgg
     4921 tgtcaaaaat aataggggcc gctgtcatca gagtaagttt aaactgagtt ctactaacta
     4981 acgagtaata tttaaatttt cagcatctcg cgcccgtgcc tctgacttct aagtccaatt
     5041 actcttcaac atccctacat gctctttctc cctgtgctcc caccccctat ttttgttatt
     5101 atcaaaaaac ttctcttaat ttctttgttt tttagcttct tttaagtcac ctctaacaat
     5161 gaaattgtgt agattcaaaa atagaattaa ttcgtaataa aaagtcgaaa aaaattgtgc
     5221 tccctccccc cattaataat aattctatcc caaaatctac acaatgttct gtgtacactt
     5281 cttatgtttt ttacttctga taaatttttt tgaaacatca tagaaaaaac cgcacacaaa
     5341 ataccttatc atatgttacg tttcagttta tgaccgcaat ttttatttct tcgcacgtct
     5401 gggcctctca tgacgtcaaa tcatgctcat cgtgaaaaag ttttggagta tttttggaat
     5461 ttttcaatca agtgaaagtt tatgaaatta attttcctgc ttttgctttt tggggtttcc
     5521 cctattgttt gtcaagattt cgaggacggc gtttttcttg ctaaaatcac aagtattgat
     5581 gagcacgatg caagaaagat cggaagaagg tttgggtttg aggctcagtg gaaggtgagt
     5641 agaagttgat aatttgaaag tggagtagtg tctatggggt ttttgcctta aatgacagaa
     5701 tacattccca atataccaaa cataactgtt tcctactagt cggccgtacg ggccctttcg
     5761 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5821 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5881 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     5941 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     6001 gccttaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     6061 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     6121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     6181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     6241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     6301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     6361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     6421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     6481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     6541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     6721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     6781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     6841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     7021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     7081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     7141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     7201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     7261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     7321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     7381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     7441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     7501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     7561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     7621 agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     7741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     7801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     7861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     7921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     7981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     8041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     8101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     8161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagct
//
