LOCUS       sequence-130-exp        8414 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 8414)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1997
REFERENCE   2  (bases 1 to 8414)
  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: L2685
FEATURES             Location/Qualifiers
     source          1..8414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             583..603
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(646..674)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1422..1442
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1575..1597)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5988..6010)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6110..6129
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6207..6225)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6293..6397
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6398..6466,6467..7258)
                     /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(6616..6635)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7429..8017
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7918..7937
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8171..8188
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8305..8326
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8341..8346,8347..8364,8365..8371)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8379..8395
                     /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     8384..8406
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(8403..8414,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(8403..8414,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tcaccaaaaa
       61 cggaacgttg agctggacgg aaatagtggt aaagtgacat gattatagtt tgaagatttc
      121 taatttcaca attagagcaa atgttgttcg gtatttattt tcaacggtat ttatactatt
      181 ttccaccttt ttctagaaca ttcgagctgc ttgttgcaaa aggagggcga ctcacattcg
      241 gtacatggaa aagtagtgta cacaataaag agacccagat acattttccg tctgcgtctc
      301 tttgcaccca ccgggagtat tttcaaacga atgcatctag gaccttctag aacattctgt
      361 aaggctgcag aatgcgggta tataaggaaa gcgggctcag aggaagccaa cacgctttgt
      421 tctagtgcat ctaaaaaact tcgaaaatcc tcatcggatc cccgggattg gccaaaggac
      481 ccaaaggtat gtttcgaatg atactaacat aacatagaac attttcagga ggacccttgg
      541 ctagaggatc cccgggtacc gagctcagaa aaaatgactg ctccaaagaa gaagcgtaag
      601 gtaccggtag aaaaaatgag taaaggagaa gaacttttca ctggagttgt cccaattctt
      661 gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga gggtgaaggt
      721 gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa actacctgtt
      781 ccatgggtaa gtttaaacat atatatacta actaaccctg attatttaaa ttttcagcca
      841 acacttgtca ctactttctg ttatggtgtt caatgcttct cgagataccc agatcatatg
      901 aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga aagaactata
      961 tttttcaaag atgacgggaa ctacaagaca cgtaagttta aacagttcgg tactaactaa
     1021 ccatacatat ttaaattttc aggtgctgaa gtcaagtttg aaggtgatac ccttgttaat
     1081 agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg acacaaattg
     1141 gaatacaact ataactcaca caatgtatac atcatggcag acaaacaaaa gaatggaatc
     1201 aaagttgtaa gtttaaacat gattttacta actaactaat ctgatttaaa ttttcagaac
     1261 ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa
     1321 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa
     1381 tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttct tgagtttgta
     1441 acagctgctg ggattacaca tggcatggat gaactataca aatgcccggt gggtgaagac
     1501 cagaaacagc acctcgaact gagccgcgat attgcccagc gtttcaacgc gctgtatggc
     1561 gagatcgatc ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     1621 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     1681 gatcgccctt cccaacagtt gcgaaggtaa gtttaaacag atccatacta actaacttgt
     1741 tctgacataa ttttcagctt gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa
     1801 gcggtgccgg aaagctggct ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc
     1861 tcaaactggc agatgcacgg ttacgatgcg cccatctaca ccaacgtgac ctatcccatt
     1921 acggtcaatc cgccgtttgt tcccacggag aatccgacgg gttgttactc gctcacattt
     1981 aatgttgatg aaagctggct acaggaaggc cagacgcgaa ttatttttga tggcgttaac
     2041 tcggcgtttc atctgtggtg caacgggcgc tgggtcggtt acggccaggt aagtttaatt
     2101 aagttgatac taactaacaa gatctgatta attttcagga cagtcgtttg ccgtctgaat
     2161 ttgacctgag cgcattttta cgcgccggag aaaaccgcct cgcggtgatg gtgctgcgct
     2221 ggagtgacgg cagttatctg gaagatcagg atatgtggcg gatgagcggc attttccgtg
     2281 acgtctcgtt gctgcataaa ccgactacac aaatcagcga tttccatgtt gccactcgct
     2341 ttaatgatga tttctcccgc gctgtactgg aggctgaagt ccaggtaagt ttaaacagga
     2401 tcttactaac taacatgcta acactgaatt ttcagatgtg cggcgagttg cgtgactacc
     2461 tacgggtaac agtttcttta tggcagggtg aaacgcaggt cgccagcggc accgcgcctt
     2521 tcggcggtga aattatcgat gagcgtggtg gttatgccga tcgcgtcaca ctacgtctga
     2581 acgtcgaaaa cccgaaactg tggagcgccg aaatcccgaa tctctatcgt gcggtggttg
     2641 aactgcacac cgccgacggc acgctgattg aagcagaagc ctgcgatgtc ggtttccgcg
     2701 aggtgcggat tgaaaatggt ctgctgctgc tgaacggcaa gccgttgctg attcgaggcg
     2761 ttaaccgtca cgagcatcat cctctgcatg gtcaggtcat ggatgagcag acgatggtgc
     2821 aggatgtaag tttaaactat tcgttactaa ctaactttaa acatttaaat tttcagatcc
     2881 tgctgatgaa gcagaacaac tttaacgccg tgcgctgttc gcattatccg aaccatccgc
     2941 tgtggtacac gctgtgcgac cgctacggcc tgtatgtggt ggatgaagcc aatattgaaa
     3001 cccacggcat ggtgccaatg aatcgtctga ccgatgatcc gcgctggcta ccggcgatga
     3061 gcgaacgcgt aacgcgaatg gtgcagcgcg atcgtaatca cccgagtgtg atcatctggt
     3121 cgctggggaa tggtaagttt aaacagttga atactaacta acggagatct ttgaaatttt
     3181 cagaatcagg ccacggcgct aatcacgacg cgctgtatcg ctggatcaaa tctgtcgatc
     3241 cttcccgccc ggtgcagtat gaaggcggcg gagccgacac cacggccacc gatattattt
     3301 gcccgatgta cgcgcgcgtg gatgaagacc agcccttccc ggctgtgccg aaatggtcca
     3361 tcaaaaaatg gctttcgcta cctggagaga cgcgcccgct gattctttgc gaggtaagtt
     3421 taaacagaac tctactaact aacacattag atcctaattt tcagtacgct cacgcgatgg
     3481 gcaacagtct tggcggtttc gctaaatact ggcaggcgtt tcgtcagtat ccccgtttac
     3541 agggcggctt cgtctgggac tgggtggatc agtcgctgat taaatatgat gaaaacggca
     3601 acccgtggtc ggcttacggc ggtgattttg gcgatacgcc gaacgatcgc cagttctgta
     3661 tgaacggtct ggtctttgcc gaccgcacgc cgcatccagc gtaagtttaa acaataacct
     3721 actaactaac gtagataatt taaattttca ggctgacgga agcaaaacac cagcagcagt
     3781 ttttccagtt ccgtttatcc gggcaaacca tcgaagtgac cagcgaatac ctgttccgtc
     3841 atagcgataa cgagctcctg cactggatgg tggcgctgga tggtaagccg ctggcaagcg
     3901 gtgaagtgcc tctggatgtc gctccacaag gtaaacagtt gattgaactg cctgaactac
     3961 cgcagccgga gagcgccggg caactctggc tcacagtacg cgtagtgcaa ccgaacgcga
     4021 ccgcatggtc agaagccggg cacatcagcg catggcagca gtggaggtaa gtttaaacaa
     4081 gatcctacta actaactcta cattgatgaa ttttcagact ggcggaaaac ctcagtgtga
     4141 cgctccccgc cgcgtcccac gccatcccgc atctgaccac cagcgaaatg gatttttgca
     4201 tcgagctggg taataagcgt tggcaattta accgccagtc aggctttctt tcacagatgt
     4261 ggattggcga taaaaaacaa ctgctgacgc cgctgcgcga tcagttcacc cgtgcaccgc
     4321 tggataacga cattggcgta agtgaagcga cccgcattga ccctaacgcc tgggtcggta
     4381 agtttaaaca aagttgtact aactaacgaa gatcttgata attttcagaa cgctggaagg
     4441 cggcgggcca ttaccaggcc gaagcagcgt tgttgcagtg cacggcagat acacttgctg
     4501 atgcggtgct gattacgacc gctcacgcgt ggcagcatca ggggaaaacc ttatttatca
     4561 gccggaaaac ctaccggatt gatggtagtg gtcaaatggc gattaccgtt gatgttgaag
     4621 tggcgagcga tacaccgcat ccggcgcgga ttggcctgaa ctgccagctg gcgcaggtag
     4681 cagagcgggt aaactggctc ggattagggc cgcaagaaaa ctatcccgac cgccttactg
     4741 ccgcctgttt tgaccgctgg gatctgccat tgtcagacat gtagtaagtt taaacttgat
     4801 agtactaact aacatgtttc atttaaattt tcagtacccc gtacgtcttc ccgagcgaaa
     4861 acggtctgcg ctgcgggacg cgcgaattga attatggccc acaccagtgg cgcggcgact
     4921 tccagttcaa catcagccgc tacagtcaac agcaactgat ggaaaccagc catcgccatc
     4981 tgctgcacgc ggaagaaggc acatggctga atatcgacgg tttccatatg gggattggtg
     5041 gcgacgactc ctggagcccg tcagtatcgg cggaattcca actgagcgcc ggtcgctacc
     5101 attaccaact tgtctggtgt caaaaataat aggggccgct gtcatcagag taagtttaaa
     5161 ctgagttcta ctaactaacg agtaatattt aaattttcag catctcgcgc ccgtgcctct
     5221 gacttctaag tccaattact cttcaacatc cctacatgct ctttctccct gtgctcccac
     5281 cccctatttt tgttattatc aaaaaacttc tcttaatttc tttgtttttt agcttctttt
     5341 aagtcacctc taacaatgaa attgtgtaga ttcaaaaata gaattaattc gtaataaaaa
     5401 gtcgaaaaaa attgtgctcc ctccccccat taataataat tctatcccaa aatctacaca
     5461 atgttctgtg tacacttctt atgtttttta cttctgataa atttttttga aacatcatag
     5521 aaaaaaccgc acacaaaata ccttatcata tgttacgttt cagtttatga ccgcaatttt
     5581 tatttcttcg cacgtctggg cctctcatga cgtcaaatca tgctcatcgt gaaaaagttt
     5641 tggagtattt ttggaatttt tcaatcaagt gaaagtttat gaaattaatt ttcctgcttt
     5701 tgctttttgg ggtttcccct attgtttgtc aagatttcga ggacggcgtt tttcttgcta
     5761 aaatcacaag tattgatgag cacgatgcaa gaaagatcgg aagaaggttt gggtttgagg
     5821 ctcagtggaa ggtgagtaga agttgataat ttgaaagtgg agtagtgtct atggggtttt
     5881 tgccttaaat gacagaatac attcccaata taccaaacat aactgtttcc tactagtcgg
     5941 ccgtacgggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     6001 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     6061 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     6121 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     6181 aaataccgca tcaggcggcc ttaagggcct cgtgatacgc ctatttttat aggttaatgt
     6241 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     6301 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6361 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     6421 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     6481 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     6541 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     6601 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     6661 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     6721 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     6781 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     6841 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     6901 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     6961 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     7021 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     7081 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     7141 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     7201 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     7261 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     7321 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     7381 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     7441 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     7501 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     7561 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     7621 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     7681 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     7741 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     7801 gagataccta cagcgtgagc attgagaaag cgccacgctt cccgaaggga gaaaggcgga
     7861 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     7921 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     7981 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     8041 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     8101 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     8161 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     8221 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     8281 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     8341 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     8401 cacaggaaac agct
//
