LOCUS       sequence-236-exp        4253 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 4253)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 4253)
  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: L4047
FEATURES             Location/Qualifiers
     source          1..4253
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             150..170
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             192..212
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             234..254
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             276..296
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(339..367)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1115..1135
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1693..1715)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1815..1834
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1912..1930)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1998..2102
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2103..2171,2172..2963)
                     /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(2321..2340)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3134..3722
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3623..3642
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3876..3893
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4010..4031
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4046..4051,4052..4069,4070..4076)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4084..4100
                     /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     4089..4111
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4108..4124
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4108..4124
                     /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 cttgcttgga
      121 gggtaccgag ctcagaaaaa atgactgctc caaagaagaa gcgtaaggta ccggcagaaa
      181 aaatgactgc tccaaagaag aagcgtaagg taccggcaga aaaaatgact gctccaaaga
      241 agaagcgtaa ggtaccggca gaaaaaatga ctgctccaaa gaagaagcgt aaggtaccgg
      301 tagaaaaaat gagtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat
      361 tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa
      421 catacggaaa acttaccctt aaatttattt gcactactgg aaaactacct gttccatggg
      481 taagtttaaa catatatata ctaactaacc ctgattattt aaattttcag ccaacacttg
      541 tcactacttt ctgttatggt gttcaatgct tctcgagata cccagatcat atgaaacggc
      601 atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaaagaact atatttttca
      661 aagatgacgg gaactacaag acacgtaagt ttaaacagtt cggtactaac taaccataca
      721 tatttaaatt ttcaggtgct gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg
      781 agttaaaagg tattgatttt aaagaagatg gaaacattct tggacacaaa ttggaataca
      841 actataactc acacaatgta tacatcatgg cagacaaaca aaagaatgga atcaaagttg
      901 taagtttaaa catgatttta ctaactaact aatctgattt aaattttcag aacttcaaaa
      961 ttagacacaa cattgaagat ggaagcgttc aactagcaga ccattatcaa caaaatactc
     1021 caattggcga tggccctgtc cttttaccag acaaccatta cctgtccaca caatctgccc
     1081 tttcgaaaga tcccaacgaa aagagagacc acatggtcct tcttgagttt gtaacagctg
     1141 ctgggattac acatggcatg gacgaactat acaaaagccc acaagctttg gaattcgcta
     1201 gccggccata caagtaatcc ggatgatcga cgccaacgtc gttgaatttt caaattttaa
     1261 atactgaata tttgtttttt ttcctattat ttatttattc tctttgtgtt ttttttcttg
     1321 ctttctaaaa aattaattca atccaaatct aaacattttt ttttctcttt ccgtctccca
     1381 attcgtattc cgctcctctc atctgaacac aatgtgcaag tttatttatc ttctcgcttt
     1441 catttcatta ggacgtgggg ggaattggtg gaagggggaa acacacaaaa ggatgatgga
     1501 aatgaaataa ggacacacaa tatgcaacaa cattcaattc agaaatatgg aggaaggttt
     1561 aaaagaaaac ataaaaatat atagaggagg aaggaaaact agtaaaaaat aagcaaagaa
     1621 attaggcgaa cgatgagaat tgtcctcgct tgggcccttt cgtctcgcgc gtttcggtga
     1681 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     1741 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     1801 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     1861 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cggccttaag ggcctcgtga
     1921 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     1981 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     2041 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2101 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     2161 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     2221 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     2281 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     2341 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     2401 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     2461 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     2521 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     2581 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     2641 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     2701 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     2761 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     2821 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     2881 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     2941 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     3001 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     3061 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3121 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3181 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3241 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     3301 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3361 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3421 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3481 tggagcgaac gacctacacc gaactgagat acctacagcg tgagcattga gaaagcgcca
     3541 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3601 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3661 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3721 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3781 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     3841 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     3901 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     3961 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     4021 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     4081 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     4141 tgtaagttta aacatgatct tactaactaa ctattctcat ttaaattttc agagcttaaa
     4201 aatggctgaa atcactcaca acgatggata cgctaacaac ttggaaatga aat
//
