LOCUS       sequence-287-exp        7470 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 7470)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 7470)
  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: L4759
FEATURES             Location/Qualifiers
     source          1..7470
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3493..3513
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3556..3584)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4332..4352
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4910..4932)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5032..5051
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5129..5147)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5215..5319
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5320..5388,5389..6180)
                     /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(5538..5557)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6351..6939
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6840..6859
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7093..7110
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7227..7248
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7263..7268,7269..7286,7287..7293)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7301..7317
                     /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     7306..7328
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7325..7341
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7325..7341
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttctgg agattgctcc agcagctctt aacagtgtct acgaccgtct tcgtgcaatt
       61 ctcatggaaa ctgaaagatc ggaaaatgca ctggatcgac gtattcagta tatgattgag
      121 actgcaatgc agattcgaaa ggacaaattt gcggtaaggt agaatatata aatagtttat
      181 tagaaaaaaa taaattagaa taatttaaat tcctactagc caatcaggcg acctttttgc
      241 gcatagttct attattgaaa aatttggaga atttctcata ttctcgctcg gaaatctgga
      301 attcgacgag atcttctggc ttctgtgcag ctgcatcgct ttgtgctccc tttctcgctt
      361 gtcttctgtg tacaccaaga accttgttga gttcatcaac tgaatctgtg actggcttgt
      421 tgctcactgg atgcactaga cgactgattc tcgagaaatc agattgagtt gcgattaggg
      481 tgacctagaa attgggaata atacgaactt ttgaaaatat tcaggaggat taaaaaaatt
      541 attctcgaca atcctacaaa tttacttatt gcaccatgtt gctccaacat ttttcattaa
      601 aagttaatga aaaaatgtag aaaatcggaa attggcaatt ttcagaccat ttttaagcat
      661 tttcaaaaaa aaattgcagc tgaaataaat gtcattttca gataaatcga gcgattttct
      721 gttgtctgac actagttttt agttttaaaa aatgttggaa gaacatggtg caataggtaa
      781 tttcatagaa tttccatgtg ttttttttca attaaccaat tatccaaatc ttccaaactc
      841 acattttgcg gagctgggct atcaagaatc tgctgcagtt ttataagacg agcatctctg
      901 atatcactga aaattaattt ttaatcaaaa cttgaatatc aactaaaccc acttattaac
      961 tttctcgatc ttctgtcgtt cggtacgatg acggtgaaga agccaattgt agtagttgat
     1021 ttggttcaag tcctttcggt gttgtacgtc agtgtcctgc aatgctattt agttataact
     1081 taggcctaag attcaattta atgaagtgat taaatttgtt ctctgaacct cttaagatga
     1141 tcttttggat tagaaacata taagacaggt ttacctatct attaaaaaac agatcaaaat
     1201 agatacgacc aaatcggata atccatgcct acctggcatc taggaacgtg ttcttagaag
     1261 atttcttacg taatcgtatg aagaaataac aatttgatcg ttggccagca aaaatagggt
     1321 tttaagtggg atagtgtttt tattagctaa ccggaaaatt ttatagtttt tttttgcaag
     1381 aaaccactga aaacccccta attgtataca ttttttggag cagcttctgg tctttttgag
     1441 caataaaatt cgataaaaca gaatttaagt gtaaattgtt cacatttagt ttctatttta
     1501 tcaaattttg ttgctcaaaa acattcgaag ctgctctaaa aaaatgcatt aaaaaagggg
     1561 ttttcagtgg tttttcacat taaaaaagct aattttaact aaaaatccat catatttcca
     1621 actttgtcac aacaataaaa tgctggtcaa aatgtgttcg aaaaaatgtt ttttttttta
     1681 atttttataa tttaaaaata gttttctttc gctgggacac atacattttt gggcgtaaat
     1741 tttcagttca aatttccatt tttacaacca taatcataaa gctacgtctg atctctctcg
     1801 cacttacctg cgcctgattc gaaagaacaa ccgtagccaa aagaacaaga agaacaagca
     1861 cgtagttgtg gtagtggacg ttcatcacgc aatactgacc aatggtcgtg gggtctcact
     1921 ttccgtacta ttgagagagg ggagactgaa gatggcaatt gaggacagtg tcttcgacgc
     1981 acgcatgcat ccataagcat aatccaggag ggatggagag aaaaatcttg tttctaagcc
     2041 cctccctttg taatacatac acatatctaa taccgaagaa tggctaattg aatggacgtc
     2101 agctgttgct gtagttgcca aggcatcatc gatgaaataa ctgaaagaaa gaattaaata
     2161 attattgcag gcgtatccgg cggtcattga agacttggac ttgattgagg aggaggatca
     2221 gatcatccat acacttaatt tggaggatgc ggttgatccg gaaaatgggc ttagtaagtg
     2281 actgaccaca cgcggggggc attaatttaa taaattgaat tccatttcag atgtgttcaa
     2341 actagatcca gaattcgaaa agaacgagga ggtttatgag gagatccgta aggaaatcat
     2401 tggaaacgcc gatatttcgg atgaggatgg tggcgacgag ttggatgatg aagaagaggg
     2461 tagtgatgtg gaagaggctc cgaagaagac tacagagatt attgataata ctgatcagaa
     2521 ttgactgctt tcagaaggta ttcattttga gttttgggcc ggcaaatctg taagttgccg
     2581 gttgccgaaa atttgctgaa tttgccggaa aaaaaaattc cggaatttat ttaaaaactt
     2641 tttgtaaaaa ttaaattaaa tttgcaactt ttcagagaag tctacctgac aatgcaatca
     2701 tctttggact accaagaagc tgctcacaaa ttgctgaaaa tgaagattcc agacagcatg
     2761 caggtcagcg atgttgcaaa gaaaaatttt cgaccaaaaa aaccaaccaa tcataaaatt
     2821 taaaaaaaaa ctccgttttt ttcttttttt ttatacgaga aaaaccaaaa aaatgtattt
     2881 ttgccaaatt ctaaaatact atccccgaaa ttttcaatat tttctctttc agaacgaact
     2941 ctgcgcgatg cttgtcgatt gttgtgctca acagcgtacc tacgagcgat tctacggaat
     3001 gctgagaaca cgttaaacac gaagatcata tttattttga gacccggatg ctctgaaaat
     3061 gtctgacata gatttaaaaa agcatatata tatttttcat tttcaacgtg aaagttttgt
     3121 gcaactttat agaatctcct attggcacat tgttttttat ttaactgagg cagtttttga
     3181 acaccttttt gaaactttga atctctttga agtatactgt cgaaaagact gacttgagcg
     3241 ttcgaaatgc cagaagaaaa ctatatttga atctcgcgct aaattgagaa atgcaaccgc
     3301 gctccactgg acaattggaa aaaaaattta ttcggaggcg acaacggtat tttcgaaatt
     3361 gattttctgt gtattttctc attttttata aattcttctt tgatttatcg ttcgtttgtg
     3421 agaaatttaa ttgtattcaa acttttttat agtaagatac cggtggtacc gagctcagaa
     3481 aaaatgacgg cacctaaaaa gaaacgcaaa gtaccggtag aaaaaatgag taaaggagaa
     3541 gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac
     3601 aaattttctg tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa
     3661 tttatttgca ctactggaaa actacctgtt ccatgggtaa gtttaaacat atatatacta
     3721 actaaccctg attatttaaa ttttcagcca acacttgtca ctactttctg ttatggtgtt
     3781 caatgcttct cgagataccc agatcatatg aaacggcatg actttttcaa gagtgccatg
     3841 cccgaaggtt atgtacagga aagaactata tttttcaaag atgacgggaa ctacaagaca
     3901 cgtaagttta aacagttcgg tactaactaa ccatacatat ttaaattttc aggtgctgaa
     3961 gtcaagtttg aaggtgatac ccttgttaat agaatcgagt taaaaggtat tgattttaaa
     4021 gaagatggaa acattcttgg acacaaattg gaatacaact ataactcaca caatgtatac
     4081 atcatggcag acaaacaaaa gaatggaatc aaagttgtaa gtttaaacat gattttacta
     4141 actaactaat ctgatttaaa ttttcagaac ttcaaaatta gacacaacat tgaagatgga
     4201 agcgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt
     4261 ttaccagaca accattacct gtccacacaa tctgcccttt cgaaagatcc caacgaaaag
     4321 agagaccaca tggtccttct tgagtttgta acagctgctg ggattacaca tggcatggac
     4381 gaactataca aaagcccaca agctttggaa ttcgctagcc ggccatacaa gtaatccgga
     4441 tgatcgacgc caacgtcgtt gaattttcaa attttaaata ctgaatattt gttttttttc
     4501 ctattattta tttattctct ttgtgttttt tttcttgctt tctaaaaaat taattcaatc
     4561 caaatctaaa catttttttt tctctttccg tctcccaatt cgtattccgc tcctctcatc
     4621 tgaacacaat gtgcaagttt atttatcttc tcgctttcat ttcattagga cgtgggggga
     4681 attggtggaa gggggaaaca cacaaaagga tgatggaaat gaaataagga cacacaatat
     4741 gcaacaacat tcaattcaga aatatggagg aaggtttaaa agaaaacata aaaatatata
     4801 gaggaggaag gaaaactagt aaaaaataag caaagaaatt aggcgaacga tgagaattgt
     4861 cctcgcttgg gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     4921 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     4981 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     5041 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     5101 gaaaataccg catcaggcgg ccttaagggc ctcgtgatac gcctattttt ataggttaat
     5161 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5221 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5281 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5341 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5401 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5461 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5521 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5581 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5641 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5701 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5761 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5821 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5881 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5941 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6001 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6061 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6121 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6181 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6241 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6301 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6361 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6421 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6481 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     6541 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6601 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6661 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6721 ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6781 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6841 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6901 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6961 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     7021 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     7081 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     7141 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     7201 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     7261 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     7321 cacacaggaa acagctatga ccatgattac gccaagctgt aagtttaaac atgatcttac
     7381 taactaacta ttctcattta aattttcaga gcttaaaaat ggctgaaatc actcacaacg
     7441 atggatacgc taacaacttg gaaatgaaat
//
