LOCUS       sequence-229-exp        6637 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 6637)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 6637)
  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: L4037
FEATURES             Location/Qualifiers
     source          1..6637
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2534..2554
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2576..2596
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2618..2638
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2660..2680
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2723..2751)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3499..3519
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4077..4099)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4199..4218
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4296..4314)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4382..4486
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4487..4555,4556..5347)
                     /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(4705..4724)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5518..6106
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6007..6026
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6260..6277
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6394..6415
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6430..6435,6436..6453,6454..6460)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6468..6484
                     /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     6473..6495
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6492..6508
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6492..6508
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgggc tgcaggtcgg ctataataag ttcttgaata aaataatttt cccgacaaaa
       61 catgagtatt tctttcgaaa ataaaagtgc aggctaatta gagattattc tgtaattaac
      121 tgcataattt gtcacgtgcc atagttttac attccactac gtcatagttc ttaaaatact
      181 aatctcctga aaatagaagt aggtgaagaa agtttaatta tcagttctaa aatgacaatt
      241 gatctttgga atatgttctg aaactaccga tcattgaaca gatgctattt gaatgatata
      301 gaattgtata tttgcaattt ctgaaacgcg ttcttaaagg cacacagatt aattcaaaag
      361 ggtctggccg caaaaaggtt tatggtggcc gattttgagt tttgtgtgtg attgcttttt
      421 cacaatcagt gttttcagga ttatgtgatg aactagatct tcaagtttcg ttacatttca
      481 tatgttttcg gaactcacga agtacatatt gggtattgtg ctcaaaaaat tcagcaatca
      541 gcttcgctcc gctgacttta gaacccaaaa aaatagtatg gccaaactga ctgtgttacg
      601 atcatttcaa tttttcaata catatttaag atttctaaga gtaagaaggt caaaaactgt
      661 tctggaatac atatatattt ttcaggttac aattagtcaa aaagtgcact gaaatatacg
      721 ttttaatttc acgaataacc caattagttc aatgtatttt tggtcaacca acgttaaagt
      781 ttggcttcca accaattatc atttctgatc aaccacaatg ttttttcttt atctgcaagt
      841 taatttttta tttttatcca gatgtttggc atatttttca attcttcact agcgcccact
      901 tcttgcactt ccggcgccct gaatctaatg catctgttgc aagaattgaa agaccaatca
      961 acacattgtt ttcttcacga gatactgaag aaaatgaata aaaacagaga aaaagagcca
     1021 tgtgattagt gacaactgtt gctaacagat accatagctt ggacttggta cgtgatggca
     1081 acgtatgggt caacaaaaat gattgcagag ggggtgcaaa acagtcaagt cgagaaaata
     1141 tgaaaaacag aaaacaaaga acagaaaaat gggtttgaga gtcagtataa tttataaaag
     1201 aaaaattgta catagaaatt aaccattttt gtagaagaag ttatttttca agcatcgtta
     1261 aaaattattc aaagcacctt atttcatatt taattttaaa catggttaaa tgaacaacac
     1321 ggtgcgcaat caggaaaact tgaaatctga aactgttgtt gtgatcttct tcgcaactgt
     1381 tcagatagca ctagtgtaat gttaagagtg cgcgaatata atggaatata atggatcaca
     1441 cctcctgcca tcaggtaaac gtctctgtta tcacatattt ccaactatta aatttttacc
     1501 ttttacagtt ttacattttt ttgaaaaaag taactttttg tcttcaaaat ccctgacgaa
     1561 aatatcaaat attttaatcg agactgcaga ggaaccgatt gatgatttgg aaaatccagc
     1621 tttacctgtg taagaactga aaagtttcat aaccctaggg tattcccagt tacattcccc
     1681 actggctaac aatagcaccc agtttttcat cacctttctt caaatttctc ggcgatttgt
     1741 taaaaacaaa atttgtgtcc cttctctgat atctctatgt ctctaaacac aagttcatcg
     1801 gaaaacgaag gagggtaggt gttggttggg ctcccgaagt gaaaatagaa gagcaagaat
     1861 agaatattag agagagagtg cagagagggc gggatagctc ccgggattcc gttttcttct
     1921 tctttatctt caacgatgat gtgtgtgcgt gttgtataga ttctgttgct cccccacaac
     1981 tcgctccgaa ggctcaatac aattcaattg atattggagg agagcctacc ggagtgggag
     2041 gataagaaga aacataagaa gaagaagaag aagaagcatg cttctggttt ttgatgctat
     2101 gaaaacggca caaaaagatg attgaggtcc cttttcaata ccttctctca tctttcaaat
     2161 cccattgaaa cctaaaactt ctcaccacgc tttaccattg ttctccaaaa acttatagca
     2221 atgtctataa cttttttatc tctgaaaagc agtgttccat ttttcttttt cctattttat
     2281 ttcaattgtt tctcacattt cgtttggatt ctttgcttgt caaccagctt cttcttccac
     2341 ttttaccgtc taattttcag ggcagggagc catcaaaccc acgaccacta gatccatcta
     2401 gaggatccta tcgattcgcg gccgctgtac acccccaaag gacccaaagg tatgtttcga
     2461 atgatactaa cataacatag aacattttca ggaggaccct tggagggtac cgagctcaga
     2521 aaaaatgact gctccaaaga agaagcgtaa ggtaccggca gaaaaaatga ctgctccaaa
     2581 gaagaagcgt aaggtaccgg cagaaaaaat gactgctcca aagaagaagc gtaaggtacc
     2641 ggcagaaaaa atgactgctc caaagaagaa gcgtaaggta ccggtagaaa aaatgagtaa
     2701 aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa
     2761 tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac
     2821 ccttaaattt atttgcacta ctggaaaact acctgttcca tgggtaagtt taaacatata
     2881 tatactaact aaccctgatt atttaaattt tcagccaaca cttgtcacta ctttctgtta
     2941 tggtgttcaa tgcttctcga gatacccaga tcatatgaaa cggcatgact ttttcaagag
     3001 tgccatgccc gaaggttatg tacaggaaag aactatattt ttcaaagatg acgggaacta
     3061 caagacacgt aagtttaaac agttcggtac taactaacca tacatattta aattttcagg
     3121 tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa aaggtattga
     3181 ttttaaagaa gatggaaaca ttcttggaca caaattggaa tacaactata actcacacaa
     3241 tgtatacatc atggcagaca aacaaaagaa tggaatcaaa gttgtaagtt taaacatgat
     3301 tttactaact aactaatctg atttaaattt tcagaacttc aaaattagac acaacattga
     3361 agatggaagc gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc
     3421 tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcga aagatcccaa
     3481 cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga ttacacatgg
     3541 catggacgaa ctatacaaaa gcccacaagc tttggaattc gctagccggc catacaagta
     3601 atccggatga tcgacgccaa cgtcgttgaa ttttcaaatt ttaaatactg aatatttgtt
     3661 ttttttccta ttatttattt attctctttg tgtttttttt cttgctttct aaaaaattaa
     3721 ttcaatccaa atctaaacat ttttttttct ctttccgtct cccaattcgt attccgctcc
     3781 tctcatctga acacaatgtg caagtttatt tatcttctcg ctttcatttc attaggacgt
     3841 ggggggaatt ggtggaaggg ggaaacacac aaaaggatga tggaaatgaa ataaggacac
     3901 acaatatgca acaacattca attcagaaat atggaggaag gtttaaaaga aaacataaaa
     3961 atatatagag gaggaaggaa aactagtaaa aaataagcaa agaaattagg cgaacgatga
     4021 gaattgtcct cgcttgggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     4081 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     4141 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     4201 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     4261 gtaaggagaa aataccgcat caggcggcct taagggcctc gtgatacgcc tatttttata
     4321 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     4381 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4441 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     4501 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4561 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     4621 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     4681 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     4741 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     4801 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     4861 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     4921 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     4981 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5041 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5101 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5161 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5221 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5281 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5341 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     5401 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5461 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5521 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5581 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5641 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     5701 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5761 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5821 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5881 caccgaactg agatacctac agcgtgagca ttgagaaagc gccacgcttc ccgaagggag
     5941 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6001 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6061 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6121 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6181 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     6241 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     6301 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     6361 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     6421 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     6481 acaatttcac acaggaaaca gctatgacca tgattacgcc aagctgtaag tttaaacatg
     6541 atcttactaa ctaactattc tcatttaaat tttcagagct taaaaatggc tgaaatcact
     6601 cacaacgatg gatacgctaa caacttggaa atgaaat
//
