LOCUS       sequence-231-exp        5537 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 5537)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 5537)
  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: L4040
FEATURES             Location/Qualifiers
     source          1..5537
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1434..1454
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1476..1496
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1518..1538
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1560..1580
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1623..1651)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2399..2419
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2977..2999)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3099..3118
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3196..3214)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3282..3386
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3387..3455,3456..4247)
                     /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(3605..3624)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4418..5006
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4907..4926
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5160..5177
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5294..5315
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5330..5335,5336..5353,5354..5360)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5368..5384
                     /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     5373..5395
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5392..5408
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5392..5408
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgaggcattt gaattggggg tggtggacag taactgtctg
       61 taataataat tactcctgac caggttgcaa ttcgagtttt gataagcata attatacctt
      121 gtacattgtg ggttttgtgc tgtggacgtt ttattgtgac atccccataa gctacaagaa
      181 caaaaatgaa atgtaaaagt attgaaaacg tcgtacattt tatatctgag tagtatcctt
      241 tgctttaaat gtccataaaa ctaattttat aatcaataaa acaacgtttg taaatcaact
      301 gagtttacaa gtagagacat tgagggatac tttcactatg ctaaagtgat aatcgaccaa
      361 ataataactt cactttggta tttattcctg tcttataaat gttatgtatg aattaaattc
      421 atatgcatat ggctcactct gacaattttt aataatcttc cagatcaata ttgactaccg
      481 atgcgggtgg tcttttgctt tgaattctgc tgaactttac accccgaaca gcaatgtgtg
      541 cttcagccta aaaaaaagta agtgtgttaa tcagtgcccc cgattcttca ttttttgccc
      601 ctctctcccg tttcgtcggc aaaagaagag aaaataaaga taagtctcaa gataggttgg
      661 taatcgctaa agtggttgtg tggataagag tagcaaaatg gcaggaagag cactttgcgc
      721 gcacacactg tactcattgt tctggataaa attctctcgt tgtttgccgt cggatgtctg
      781 cctctctgca ttgagccggc ttcttcacta tctttagtta acctaaaatg ccgtttcttt
      841 tctcgtatcc ccactatccc gttgaggttc tctgctctct tcgctcccta ccgccagcga
      901 gcaactatcc gtgggggcgc cttgctcgga agatgggggg gaagaaagaa gatttttgct
      961 atttgcactt gagaaagaga cttttcctgc gtcgatggtt agagaacagt gtgcagacac
     1021 ttttcagcta cctagaatta caattggata tccccgcctc ccaatccacc cacccaggga
     1081 aaaagaaggg ctcgccgaaa atcaaagtta tctccaggct cgcgcatccc accgagcggt
     1141 tgacttctct ccaccacttt tcattttaac cctcggggta cgggattggc caaaggaccc
     1201 aaaggtatgt ttcgaatgat actaacataa catagaacat tttcaggagg acccttggct
     1261 agcgtcgacg gtaccggtca gatatcactc gaggagctcc cgagatccta tcgattcgcg
     1321 gccgctgtac acccccaaag gacccaaagg tatgtttcga atgatactaa cataacatag
     1381 aacattttca ggaggaccct tggagggtac cgagctcaga aaaaatgact gctccaaaga
     1441 agaagcgtaa ggtaccggca gaaaaaatga ctgctccaaa gaagaagcgt aaggtaccgg
     1501 cagaaaaaat gactgctcca aagaagaagc gtaaggtacc ggcagaaaaa atgactgctc
     1561 caaagaagaa gcgtaaggta ccggtagaaa aaatgagtaa aggagaagaa cttttcactg
     1621 gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca
     1681 gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta
     1741 ctggaaaact acctgttcca tgggtaagtt taaacatata tatactaact aaccctgatt
     1801 atttaaattt tcagccaaca cttgtcacta ctttctgtta tggtgttcaa tgcttctcga
     1861 gatacccaga tcatatgaaa cggcatgact ttttcaagag tgccatgccc gaaggttatg
     1921 tacaggaaag aactatattt ttcaaagatg acgggaacta caagacacgt aagtttaaac
     1981 agttcggtac taactaacca tacatattta aattttcagg tgctgaagtc aagtttgaag
     2041 gtgataccct tgttaataga atcgagttaa aaggtattga ttttaaagaa gatggaaaca
     2101 ttcttggaca caaattggaa tacaactata actcacacaa tgtatacatc atggcagaca
     2161 aacaaaagaa tggaatcaaa gttgtaagtt taaacatgat tttactaact aactaatctg
     2221 atttaaattt tcagaacttc aaaattagac acaacattga agatggaagc gttcaactag
     2281 cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc
     2341 attacctgtc cacacaatct gccctttcga aagatcccaa cgaaaagaga gaccacatgg
     2401 tccttcttga gtttgtaaca gctgctggga ttacacatgg catggacgaa ctatacaaaa
     2461 gcccacaagc tttggaattc gctagccggc catacaagta atccggatga tcgacgccaa
     2521 cgtcgttgaa ttttcaaatt ttaaatactg aatatttgtt ttttttccta ttatttattt
     2581 attctctttg tgtttttttt cttgctttct aaaaaattaa ttcaatccaa atctaaacat
     2641 ttttttttct ctttccgtct cccaattcgt attccgctcc tctcatctga acacaatgtg
     2701 caagtttatt tatcttctcg ctttcatttc attaggacgt ggggggaatt ggtggaaggg
     2761 ggaaacacac aaaaggatga tggaaatgaa ataaggacac acaatatgca acaacattca
     2821 attcagaaat atggaggaag gtttaaaaga aaacataaaa atatatagag gaggaaggaa
     2881 aactagtaaa aaataagcaa agaaattagg cgaacgatga gaattgtcct cgcttgggcc
     2941 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     3001 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     3061 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     3121 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     3181 caggcggcct taagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     3241 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3301 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3361 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3421 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3481 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     3541 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     3601 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3661 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     3721 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3781 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     3841 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3901 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     3961 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     4021 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4081 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4141 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4201 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     4261 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4321 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4381 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4441 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4501 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4561 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4621 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4681 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4741 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4801 agcgtgagca ttgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4861 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4921 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4981 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5041 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5101 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     5161 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     5221 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     5281 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     5341 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     5401 gctatgacca tgattacgcc aagctgtaag tttaaacatg atcttactaa ctaactattc
     5461 tcatttaaat tttcagagct taaaaatggc tgaaatcact cacaacgatg gatacgctaa
     5521 caacttggaa atgaaat
//
