LOCUS       sequence-274-exp       10324 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 10324)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 10324)
  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: L4660
FEATURES             Location/Qualifiers
     source          1..10324
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2493..2513
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2556..2584)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3332..3352
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3485..3507)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7898..7920)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8020..8039
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(8117..8135)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8203..8307
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8308..8376,8377..9168)
                     /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(8526..8545)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9339..9927
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9828..9847
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10081..10098
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10215..10236
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10251..10256,10257..10274,10275..10281)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10289..10305
                     /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     10294..10316
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(10313..10324,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(10313..10324,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgggctgc aggtcggcta taataagttc ttgaataaaa
       61 taattttccc gacaaaacat gagtatttct ttcgaaaata aaagtgcagg ctaattagag
      121 attattctgt aattaactgc ataatttgtc acgtgccata gttttacatt ccactacgtc
      181 atagttctta aaatactaat ctcctgaaaa tagaagtagg tgaagaaagt ttaattatca
      241 gttctaaaat gacaattgat ctttggaata tgttctgaaa ctaccgatca ttgaacagat
      301 gctatttgaa tgatatagaa ttgtatattt gcaatttctg aaacgcgttc ttaaaggcac
      361 acagattaat tcaaaagggt ctggccgcaa aaaggtttat ggtggccgat tttgagtttt
      421 gtgtgtgatt gctttttcac aatcagtgtt ttcaggatta tgtgatgaac tagatcttca
      481 agtttcgtta catttcatat gttttcggaa ctcacgaagt acatattggg tattgtgctc
      541 aaaaaattca gcaatcagct tcgctccgct gactttagaa cccaaaaaaa tagtatggcc
      601 aaactgactg tgttacgatc atttcaattt ttcaatacat atttaagatt tctaagagta
      661 agaaggtcaa aaactgttct ggaatacata tatatttttc aggttacaat tagtcaaaaa
      721 gtgcactgaa atatacgttt taatttcacg aataacccaa ttagttcaat gtatttttgg
      781 tcaaccaacg ttaaagtttg gcttccaacc aattatcatt tctgatcaac cacaatgttt
      841 tttctttatc tgcaagttaa ttttttattt ttatccagat gtttggcata tttttcaatt
      901 cttcactagc gcccacttct tgcacttccg gcgccctgaa tctaatgcat ctgttgcaag
      961 aattgaaaga ccaatcaaca cattgttttc ttcacgagat actgaagaaa atgaataaaa
     1021 acagagaaaa agagccatgt gattagtgac aactgttgct aacagatacc atagcttgga
     1081 cttggtacgt gatggcaacg tatgggtcaa caaaaatgat tgcagagggg gtgcaaaaca
     1141 gtcaagtcga gaaaatatga aaaacagaaa acaaagaaca gaaaaatggg tttgagagtc
     1201 agtataattt ataaaagaaa aattgtacat agaaattaac catttttgta gaagaagtta
     1261 tttttcaagc atcgttaaaa attattcaaa gcaccttatt tcatatttaa ttttaaacat
     1321 ggttaaatga acaacacggt gcgcaatcag gaaaacttga aatctgaaac tgttgttgtg
     1381 atcttcttcg caactgttca gatagcacta gtgtaatgtt aagagtgcgc gaatataatg
     1441 gaatataatg gatcacacct cctgccatca ggtaaacgtc tctgttatca catatttcca
     1501 actattaaat ttttaccttt tacagtttta catttttttg aaaaaagtaa ctttttgtct
     1561 tcaaaatccc tgacgaaaat atcaaatatt ttaatcgaga ctgcagagga accgattgat
     1621 gatttggaaa atccagcttt acctgtgtaa gaactgaaaa gtttcataac cctagggtat
     1681 tcccagttac attccccact ggctaacaat agcacccagt ttttcatcac ctttcttcaa
     1741 atttctcggc gatttgttaa aaacaaaatt tgtgtccctt ctctgatatc tctatgtctc
     1801 taaacacaag ttcatcggaa aacgaaggag ggtaggtgtt ggttgggctc ccgaagtgaa
     1861 aatagaagag caagaataga atattagaga gagagtgcag agagggcggg atagctcccg
     1921 ggattccgtt ttcttcttct ttatcttcaa cgatgatgtg tgtgcgtgtt gtatagattc
     1981 tgttgctccc ccacaactcg ctccgaaggc tcaatacaat tcaattgata ttggaggaga
     2041 gcctaccgga gtgggaggat aagaagaaac ataagaagaa gaagaagaag aagcatgctt
     2101 ctggtttttg atgctatgaa aacggcacaa aaagatgatt gaggtccctt ttcaatacct
     2161 tctctcatct ttcaaatccc attgaaacct aaaacttctc accacgcttt accattgttc
     2221 tccaaaaact tatagcaatg tctataactt ttttatctct gaaaagcagt gttccatttt
     2281 tctttttcct attttatttc aattgtttct cacatttcgt ttggattctt tgcttgtcaa
     2341 ccagcttctt cttccacttt taccgtctaa ttttcagggc agggagccat caaacccacg
     2401 accactagat ccatctagag gatccccggg attggccaaa ggacccaaag gaggaccctt
     2461 ggagggtacc gagctcagaa aaaatgactg ctccaaagaa gaagcgtaag gtaccggtag
     2521 aaaaaatgag taaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag
     2581 atggtgatgt taatgggcac aaattttctg tcagtggaga gggtgaaggt gatgcaacat
     2641 acggaaaact tacccttaaa tttatttgca ctactggaaa actacctgtt ccatgggtaa
     2701 gtttaaacat atatatacta actaaccctg attatttaaa ttttcagcca acacttgtca
     2761 ctactttctc atggggcgtt caatgcttct cgagataccc agatcatatg aaacagcatg
     2821 actttttcaa gagtgccatg cccgaaggtt atgtacagga aagaactata tttttcaaag
     2881 atgacgggaa ctacaagaca cgtaagttta aacagttcgg tactaactaa ccatacatat
     2941 ttaaattttc aggtgctgaa gtcaagtttg aaggtgatac ccttgttaat agaatcgagt
     3001 taaaaggtat tgattttaaa gaagatggaa acattcttgg acacaaattg gaatacaact
     3061 atatttcaca caacgtatac atcactgccg acaaacaaaa gaatgggatc aaagcggtaa
     3121 gtttaaacat gattttacta actaactaat ctgatttaaa ttttcagaac ttcaaaatta
     3181 gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa aatactccaa
     3241 ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa tctgcccttt
     3301 cgaaagatcc caacgaaaag agagaccaca tggtccttct tgagtttgta acagctgctg
     3361 ggattacaca tggcatggat gaactataca aatgcccggt gggtgaagac cagaaacagc
     3421 acctcgaact gagccgcgat attgcccagc gtttcaacgc gctgtatggc gagatcgatc
     3481 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     3541 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     3601 cccaacagtt gcgaaggtaa gtttaaacag atccatacta actaacttgt tctgacataa
     3661 ttttcagctt gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa gcggtgccgg
     3721 aaagctggct ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc tcaaactggc
     3781 agatgcacgg ttacgatgcg cccatctaca ccaacgtgac ctatcccatt acggtcaatc
     3841 cgccgtttgt tcccacggag aatccgacgg gttgttactc gctcacattt aatgttgatg
     3901 aaagctggct acaggaaggc cagacgcgaa ttatttttga tggcgttaac tcggcgtttc
     3961 atctgtggtg caacgggcgc tgggtcggtt acggccaggt aagtttaatt aagttgatac
     4021 taactaacaa gatctgatta attttcagga cagtcgtttg ccgtctgaat ttgacctgag
     4081 cgcattttta cgcgccggag aaaaccgcct cgcggtgatg gtgctgcgct ggagtgacgg
     4141 cagttatctg gaagatcagg atatgtggcg gatgagcggc attttccgtg acgtctcgtt
     4201 gctgcataaa ccgactacac aaatcagcga tttccatgtt gccactcgct ttaatgatga
     4261 tttctcccgc gctgtactgg aggctgaagt ccaggtaagt ttaaacagga tcttactaac
     4321 taacatgcta acactgaatt ttcagatgtg cggcgagttg cgtgactacc tacgggtaac
     4381 agtttcttta tggcagggtg aaacgcaggt cgccagcggc accgcgcctt tcggcggtga
     4441 aattatcgat gagcgtggtg gttatgccga tcgcgtcaca ctacgtctga acgtcgaaaa
     4501 cccgaaactg tggagcgccg aaatcccgaa tctctatcgt gcggtggttg aactgcacac
     4561 cgccgacggc acgctgattg aagcagaagc ctgcgatgtc ggtttccgcg aggtgcggat
     4621 tgaaaatggt ctgctgctgc tgaacggcaa gccgttgctg attcgaggcg ttaaccgtca
     4681 cgagcatcat cctctgcatg gtcaggtcat ggatgagcag acgatggtgc aggatgtaag
     4741 tttaaactat tcgttactaa ctaactttaa acatttaaat tttcagatcc tgctgatgaa
     4801 gcagaacaac tttaacgccg tgcgctgttc gcattatccg aaccatccgc tgtggtacac
     4861 gctgtgcgac cgctacggcc tgtatgtggt ggatgaagcc aatattgaaa cccacggcat
     4921 ggtgccaatg aatcgtctga ccgatgatcc gcgctggcta ccggcgatga gcgaacgcgt
     4981 aacgcgaatg gtgcagcgcg atcgtaatca cccgagtgtg atcatctggt cgctggggaa
     5041 tggtaagttt aaacagttga atactaacta acggagatct ttgaaatttt cagaatcagg
     5101 ccacggcgct aatcacgacg cgctgtatcg ctggatcaaa tctgtcgatc cttcccgccc
     5161 ggtgcagtat gaaggcggcg gagccgacac cacggccacc gatattattt gcccgatgta
     5221 cgcgcgcgtg gatgaagacc agcccttccc ggctgtgccg aaatggtcca tcaaaaaatg
     5281 gctttcgcta cctggagaga cgcgcccgct gattctttgc gaggtaagtt taaacagaac
     5341 tctactaact aacacattag atcctaattt tcagtacgct cacgcgatgg gcaacagtct
     5401 tggcggtttc gctaaatact ggcaggcgtt tcgtcagtat ccccgtttac agggcggctt
     5461 cgtctgggac tgggtggatc agtcgctgat taaatatgat gaaaacggca acccgtggtc
     5521 ggcttacggc ggtgattttg gcgatacgcc gaacgatcgc cagttctgta tgaacggtct
     5581 ggtctttgcc gaccgcacgc cgcatccagc gtaagtttaa acaataacct actaactaac
     5641 gtagataatt taaattttca ggctgacgga agcaaaacac cagcagcagt ttttccagtt
     5701 ccgtttatcc gggcaaacca tcgaagtgac cagcgaatac ctgttccgtc atagcgataa
     5761 cgagctcctg cactggatgg tggcgctgga tggtaagccg ctggcaagcg gtgaagtgcc
     5821 tctggatgtc gctccacaag gtaaacagtt gattgaactg cctgaactac cgcagccgga
     5881 gagcgccggg caactctggc tcacagtacg cgtagtgcaa ccgaacgcga ccgcatggtc
     5941 agaagccggg cacatcagcg catggcagca gtggaggtaa gtttaaacaa gatcctacta
     6001 actaactcta cattgatgaa ttttcagact ggcggaaaac ctcagtgtga cgctccccgc
     6061 cgcgtcccac gccatcccgc atctgaccac cagcgaaatg gatttttgca tcgagctggg
     6121 taataagcgt tggcaattta accgccagtc aggctttctt tcacagatgt ggattggcga
     6181 taaaaaacaa ctgctgacgc cgctgcgcga tcagttcacc cgtgcaccgc tggataacga
     6241 cattggcgta agtgaagcga cccgcattga ccctaacgcc tgggtcggta agtttaaaca
     6301 aagttgtact aactaacgaa gatcttgata attttcagaa cgctggaagg cggcgggcca
     6361 ttaccaggcc gaagcagcgt tgttgcagtg cacggcagat acacttgctg atgcggtgct
     6421 gattacgacc gctcacgcgt ggcagcatca ggggaaaacc ttatttatca gccggaaaac
     6481 ctaccggatt gatggtagtg gtcaaatggc gattaccgtt gatgttgaag tggcgagcga
     6541 tacaccgcat ccggcgcgga ttggcctgaa ctgccagctg gcgcaggtag cagagcgggt
     6601 aaactggctc ggattagggc cgcaagaaaa ctatcccgac cgccttactg ccgcctgttt
     6661 tgaccgctgg gatctgccat tgtcagacat gtagtaagtt taaacttgat agtactaact
     6721 aacatgtttc atttaaattt tcagtacccc gtacgtcttc ccgagcgaaa acggtctgcg
     6781 ctgcgggacg cgcgaattga attatggccc acaccagtgg cgcggcgact tccagttcaa
     6841 catcagccgc tacagtcaac agcaactgat ggaaaccagc catcgccatc tgctgcacgc
     6901 ggaagaaggc acatggctga atatcgacgg tttccatatg gggattggtg gcgacgactc
     6961 ctggagcccg tcagtatcgg cggaattcca actgagcgcc ggtcgctacc attaccaact
     7021 tgtctggtgt caaaaataat aggggccgct gtcatcagag taagtttaaa ctgagttcta
     7081 ctaactaacg agtaatattt aaattttcag catctcgcgc ccgtgcctct gacttctaag
     7141 tccaattact cttcaacatc cctacatgct ctttctccct gtgctcccac cccctatttt
     7201 tgttattatc aaaaaacttc tcttaatttc tttgtttttt agcttctttt aagtcacctc
     7261 taacaatgaa attgtgtaga ttcaaaaata gaattaattc gtaataaaaa gtcgaaaaaa
     7321 attgtgctcc ctccccccat taataataat tctatcccaa aatctacaca atgttctgtg
     7381 tacacttctt atgtttttta cttctgataa atttttttga aacatcatag aaaaaaccgc
     7441 acacaaaata ccttatcata tgttacgttt cagtttatga ccgcaatttt tatttcttcg
     7501 cacgtctggg cctctcatga cgtcaaatca tgctcatcgt gaaaaagttt tggagtattt
     7561 ttggaatttt tcaatcaagt gaaagtttat gaaattaatt ttcctgcttt tgctttttgg
     7621 ggtttcccct attgtttgtc aagatttcga ggacggcgtt tttcttgcta aaatcacaag
     7681 tattgatgag cacgatgcaa gaaagatcgg aagaaggttt gggtttgagg ctcagtggaa
     7741 ggtgagtaga agttgataat ttgaaagtgg agtagtgtct atggggtttt tgccttaaat
     7801 gacagaatac attcccaata taccaaacat aactgtttcc tactagtcgg ccgtacgggc
     7861 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     7921 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     7981 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     8041 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     8101 tcaggcggcc ttaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     8161 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     8221 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     8281 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     8341 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     8401 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     8461 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     8521 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     8581 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     8641 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     8701 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     8761 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     8821 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     8881 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     8941 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     9001 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     9061 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     9121 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     9181 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     9241 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     9301 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     9361 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     9421 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     9481 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     9541 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     9601 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     9661 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     9721 cagcgtgagc attgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     9781 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     9841 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     9901 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     9961 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
    10021 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
    10081 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
    10141 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
    10201 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
    10261 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
    10321 agct
//
