LOCUS       sequence-240-exp        4249 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 4249)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 4249)
  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: L4052
FEATURES             Location/Qualifiers
     source          1..4249
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             146..166
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             188..208
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             230..250
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             272..292
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(335..363)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1111..1131
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1689..1711)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1811..1830
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1908..1926)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1994..2098
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2099..2167,2168..2959)
                     /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(2317..2336)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3130..3718
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3619..3638
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3872..3889
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4006..4027
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4042..4047,4048..4065,4066..4072)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4080..4096
                     /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     4085..4107
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4104..4120
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4104..4120
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ggatccccgg gattggccaa aggacccaaa
       61 ggtatgtttc gaatgatact aacataacat agaacatttt caggaggacc cttggagggt
      121 accgagctca gaaaaaatga ctgctccaaa gaagaagcgt aaggtaccgg cagaaaaaat
      181 gactgctcca aagaagaagc gtaaggtacc ggcagaaaaa atgactgctc caaagaagaa
      241 gcgtaaggta ccggcagaaa aaatgactgc tccaaagaag aagcgtaagg taccggtaga
      301 aaaaatgagt aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga
      361 tggtgatgtt aatgggcaca aattttctgt cagtggagag ggtgaaggtg atgcaacata
      421 cggaaaactt acccttaaat ttatttgcac tactggaaaa ctacctgttc catgggtaag
      481 tttaaacata tatatactaa ctaaccctga ttatttaaat tttcagccaa cacttgtcac
      541 tactttctgt tatggtgttc aatgcttctc gagataccca gatcatatga aacggcatga
      601 ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa agaactatat ttttcaaaga
      661 tgacgggaac tacaagacac gtaagtttaa acagttcggt actaactaac catacatatt
      721 taaattttca ggtgctgaag tcaagtttga aggtgatacc cttgttaata gaatcgagtt
      781 aaaaggtatt gattttaaag aagatggaaa cattcttgga cacaaattgg aatacaacta
      841 taactcacac aatgtataca tcatggcaga caaacaaaag aatggaatca aagttgtaag
      901 tttaaacatg attttactaa ctaactaatc tgatttaaat tttcagaact tcaaaattag
      961 acacaacatt gaagatggaa gcgttcaact agcagaccat tatcaacaaa atactccaat
     1021 tggcgatggc cctgtccttt taccagacaa ccattacctg tccacacaat ctgccctttc
     1081 gaaagatccc aacgaaaaga gagaccacat ggtccttctt gagtttgtaa cagctgctgg
     1141 gattacacat ggcatggacg aactatacaa aagcccacaa gctttggaat tcgctagccg
     1201 gccatacaag taatccggat gatcgacgcc aacgtcgttg aattttcaaa ttttaaatac
     1261 tgaatatttg ttttttttcc tattatttat ttattctctt tgtgtttttt ttcttgcttt
     1321 ctaaaaaatt aattcaatcc aaatctaaac attttttttt ctctttccgt ctcccaattc
     1381 gtattccgct cctctcatct gaacacaatg tgcaagttta tttatcttct cgctttcatt
     1441 tcattaggac gtggggggaa ttggtggaag ggggaaacac acaaaaggat gatggaaatg
     1501 aaataaggac acacaatatg caacaacatt caattcagaa atatggagga aggtttaaaa
     1561 gaaaacataa aaatatatag aggaggaagg aaaactagta aaaaataagc aaagaaatta
     1621 ggcgaacgat gagaattgtc ctcgcttggg ccctttcgtc tcgcgcgttt cggtgatgac
     1681 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     1741 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     1801 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     1861 ccgcacagat gcgtaaggag aaaataccgc atcaggcggc cttaagggcc tcgtgatacg
     1921 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     1981 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     2041 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     2101 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     2161 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     2221 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     2281 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     2341 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     2401 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     2461 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     2521 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     2581 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     2641 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     2701 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     2761 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     2821 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     2881 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     2941 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     3001 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     3061 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3121 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3181 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3241 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     3301 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3361 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3421 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3481 gcgaacgacc tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct
     3541 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3601 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3661 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     3721 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     3781 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     3841 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     3901 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     3961 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     4021 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     4081 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagctgta
     4141 agtttaaaca tgatcttact aactaactat tctcatttaa attttcagag cttaaaaatg
     4201 gctgaaatca ctcacaacga tggatacgct aacaacttgg aaatgaaat
//
