LOCUS       sequence-271-exp        4693 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 4693)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 4693)
  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: L4632
FEATURES             Location/Qualifiers
     source          1..4693
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             10..30
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(73..101)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     849..869
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1891..1913)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2013..2032
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2110..2128)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2196..2300
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2301..2369,2370..3161)
                     /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(2519..2538)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3332..3920
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3821..3840
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4074..4091
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4208..4229
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4244..4249,4250..4267,4268..4274)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4282..4298
                     /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     4287..4309
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4306..4322
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4306..4322
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgactgctc caaagaagaa gcgtaaggta ccggtagaaa aaatgagtaa aggagaagaa
       61 cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa
      121 ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt
      181 atttgcacta ctggaaaact acctgttcca tgggtaagtt taaacatata tatactaact
      241 aaccctgatt atttaaattt tcagccaaca cttgtcacta ctttcggtta tggtcttcaa
      301 tgcttcgcca gatacccaga tcatatgaaa cagcatgact ttttcaagag tgccatgccc
      361 gaaggttatg tacaggaaag aactatattt ttcaaagatg acgggaacta caagacacgt
      421 aagtttaaac agttcggtac taactaacca tacatattta aattttcagg tgctgaagtc
      481 aagtttgaag gtgataccct tgttaataga atcgagttaa aaggtattga ttttaaagaa
      541 gatggaaaca ttcttggaca caaattggaa tacaactata actcacacaa tgtatacatc
      601 atggcagaca aacaaaagaa tggaatcaaa gttgtaagtt taaacatgat tttactaact
      661 aactaatctg atttaaattt tcagaacttc aaaattagac acaacattga agatggaagc
      721 gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta
      781 ccagacaacc attacctgtc cacacaatct gccctttcga aagatcccaa cgaaaagaga
      841 gaccacatgg tccttcttga gtttgtaaca gctgctggga ttacacatgg catggatgaa
      901 ctatacaaat agcattcgta gaattgtaag tttaaacttt ctcatactaa ctaacgcgct
      961 ctatttaaat tttcagaatt ccaactgagc gccggtcgct accattacca acttgtctgg
     1021 tgtcaaaaat aataggggcc gctgtcatca gagtaagttt aaactgagtt ctactaacta
     1081 acgagtaata tttaaatttt cagcatctcg cgcccgtgcc tctgacttct aagtccaatt
     1141 actcttcaac atccctacat gctctttctc cctgtgctcc caccccctat ttttgttatt
     1201 atcaaaaaac ttctcttaat ttctttgttt tttagcttct tttaagtcac ctctaacaat
     1261 gaaattgtgt agattcaaaa atagaattaa ttcgtaataa aaagtcgaaa aaaattgtgc
     1321 tccctccccc cattaataat aattctatcc caaaatctac acaatgttct gtgtacactt
     1381 cttatgtttt ttacttctga taaatttttt tgaaacatca tagaaaaaac cgcacacaaa
     1441 ataccttatc atatgttacg tttcagttta tgaccgcaat ttttatttct tcgcacgtct
     1501 gggcctctca tgacgtcaaa tcatgctcat cgtgaaaaag ttttggagta tttttggaat
     1561 ttttcaatca agtgaaagtt tatgaaatta attttcctgc ttttgctttt tggggtttcc
     1621 cctattgttt gtcaagattt cgaggacggc gtttttcttg ctaaaatcac aagtattgat
     1681 gagcacgatg caagaaagat cggaagaagg tttgggtttg aggctcagtg gaaggtgagt
     1741 agaagttgat aatttgaaag tggagtagtg tctatggggt ttttgcctta aatgacagaa
     1801 tacattccca atataccaaa cataactgtt tcctactagt cggccgtacg ggccctttcg
     1861 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     1921 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     1981 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     2041 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     2101 gccttaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     2161 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     2221 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     2281 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     2341 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     2401 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     2461 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     2521 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     2581 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     2641 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     2701 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     2761 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     2821 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     2881 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     2941 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     3001 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     3061 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     3121 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     3181 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     3241 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3301 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3361 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3421 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     3481 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3541 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3601 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3661 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3721 agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3781 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3841 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3901 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3961 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     4021 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     4081 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     4141 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     4201 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     4261 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     4321 accatgatta cgccaagctt gcatgcctgc aggtcgactc tagaggatcc ccaattcttg
     4381 agacttccac tatcttctct ggcaccctcc actacgcgta ggcgtctctc gcttcgtgta
     4441 ttcccgggaa gccgggttcc cgtctctccc gccgctgccg ctgccgcaca cagctttaca
     4501 cctcgtagaa tccccaaaga ggggcgtggc ttgcgggtgc caacatcctc ctgccgagga
     4561 agaagcaggc actcatcact cgcatcatca acctcgggat tggccaaagg acccaaaggt
     4621 atgtttcgaa tgatactaac ataacataga acattttcag gaggaccctt ggagggtacc
     4681 gagctcagaa aaa
//
