LOCUS       sequence-226-exp        4711 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 4711)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 4711)
  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: L4020
FEATURES             Location/Qualifiers
     source          1..4711
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             144..164
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             186..206
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             228..248
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             270..290
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(333..361)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1109..1129
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2151..2173)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2273..2292
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2370..2388)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2456..2560
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2561..2629,2630..3421)
                     /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(2779..2798)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3592..4180
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4081..4100
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4334..4351
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4468..4489
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4504..4509,4510..4527,4528..4534)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4542..4558
                     /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     4547..4569
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4566..4582
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4566..4582
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactagagg atccccggga ttggccaaag gacccaaagg
       61 tatgtttcga atgatactaa cataacatag aacattttca ggaggaccct tggagggtac
      121 cgagctcaga aaaaatgact gctccaaaga agaagcgtaa ggtaccggca gaaaaaatga
      181 ctgctccaaa gaagaagcgt aaggtaccgg cagaaaaaat gactgctcca aagaagaagc
      241 gtaaggtacc ggcagaaaaa atgactgctc caaagaagaa gcgtaaggta ccggtagaaa
      301 aaatgagtaa aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg
      361 gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg
      421 gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcca tgggtaagtt
      481 taaacatata tatactaact aaccctgatt atttaaattt tcagccaaca cttgtcacta
      541 ctttctgtta tggtgttcaa tgcttctcga gatacccaga tcatatgaaa cggcatgact
      601 ttttcaagag tgccatgccc gaaggttatg tacaggaaag aactatattt ttcaaagatg
      661 acgggaacta caagacacgt aagtttaaac agttcggtac taactaacca tacatattta
      721 aattttcagg tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa
      781 aaggtattga ttttaaagaa gatggaaaca ttcttggaca caaattggaa tacaactata
      841 actcacacaa tgtatacatc atggcagaca aacaaaagaa tggaatcaaa gttgtaagtt
      901 taaacatgat tttactaact aactaatctg atttaaattt tcagaacttc aaaattagac
      961 acaacattga agatggaagc gttcaactag cagaccatta tcaacaaaat actccaattg
     1021 gcgatggccc tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcga
     1081 aagatcccaa cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga
     1141 ttacacatgg catggatgaa ctatacaaat agcattcgta gaattgtaag tttaaacttt
     1201 ctcatactaa ctaacgcgct ctatttaaat tttcagaatt ccaactgagc gccggtcgct
     1261 accattacca acttgtctgg tgtcaaaaat aataggggcc gctgtcatca gagtaagttt
     1321 aaactgagtt ctactaacta acgagtaata tttaaatttt cagcatctcg cgcccgtgcc
     1381 tctgacttct aagtccaatt actcttcaac atccctacat gctctttctc cctgtgctcc
     1441 caccccctat ttttgttatt atcaaaaaac ttctcttaat ttctttgttt tttagcttct
     1501 tttaagtcac ctctaacaat gaaattgtgt agattcaaaa atagaattaa ttcgtaataa
     1561 aaagtcgaaa aaaattgtgc tccctccccc cattaataat aattctatcc caaaatctac
     1621 acaatgttct gtgtacactt cttatgtttt ttacttctga taaatttttt tgaaacatca
     1681 tagaaaaaac cgcacacaaa ataccttatc atatgttacg tttcagttta tgaccgcaat
     1741 ttttatttct tcgcacgtct gggcctctca tgacgtcaaa tcatgctcat cgtgaaaaag
     1801 ttttggagta tttttggaat ttttcaatca agtgaaagtt tatgaaatta attttcctgc
     1861 ttttgctttt tggggtttcc cctattgttt gtcaagattt cgaggacggc gtttttcttg
     1921 ctaaaatcac aagtattgat gagcacgatg caagaaagat cggaagaagg tttgggtttg
     1981 aggctcagtg gaaggtgagt agaagttgat aatttgaaag tggagtagtg tctatggggt
     2041 ttttgcctta aatgacagaa tacattccca atataccaaa cataactgtt tcctactagt
     2101 cggccgtacg ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     2161 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     2221 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     2281 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     2341 agaaaatacc gcatcaggcg gccttaaggg cctcgtgata cgcctatttt tataggttaa
     2401 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     2461 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2521 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     2581 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     2641 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     2701 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     2761 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     2821 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     2881 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     2941 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3001 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3061 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     3121 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     3181 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     3241 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     3301 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     3361 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     3421 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     3481 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     3541 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3601 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3661 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3721 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3781 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3841 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3901 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3961 actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc
     4021 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4081 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4141 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4201 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     4261 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     4321 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     4381 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     4441 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     4501 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     4561 tcacacagga aacagctatg accatgatta cgccaagctg taagtttaaa catgatctta
     4621 ctaactaact attctcattt aaattttcag agcttaaaaa tggctgaaat cactcacaac
     4681 gatggatacg ctaacaactt ggaaatgaaa t
//
