LOCUS       sequence-267-exp        4693 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 4693)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 4693)
  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: L4458
FEATURES             Location/Qualifiers
     source          1..4693
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             386..406
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(449..477)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1225..1245
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2267..2289)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2389..2408
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2486..2504)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2572..2676
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2677..2745,2746..3537)
                     /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(2895..2914)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3708..4296
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4197..4216
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4450..4467
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4584..4605
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4620..4625,4626..4643,4644..4650)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4658..4674
                     /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     4663..4685
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(4682..4693,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(4682..4693,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccaattc
       61 ttgagacttc cactatcttc tctggcaccc tccactacgc gtaggcgtct ctcgcttcgt
      121 gtattcccgg gaagccgggt tcccgtctct cccgccgctg ccgctgccgc acacagcttt
      181 acacctcgta gaatccccaa agaggggcgt ggcttgcggg tgccaacatc ctcctgccga
      241 ggaagaagca ggcactcatc actcgcatca tcaacctcgg gattggccaa aggacccaaa
      301 ggtatgtttc gaatgatact aacataacat agaacatttt caggaggacc cttggagggt
      361 accgagctca gaaaaaatga ctgctccaaa gaagaagcgt aaggtaccgg tagaaaaaat
      421 gagtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga
      481 tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa
      541 acttaccctt aaatttattt gcactactgg aaaactacct gttccatggg taagtttaaa
      601 catatatata ctaactaacc ctgattattt aaattttcag ccaacacttg tcactacttt
      661 ctgttatggt gttcaatgct tctcgagata cccagatcat atgaaacggc atgacttttt
      721 caagagtgcc atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg
      781 gaactacaag acacgtaagt ttaaacagtt cggtactaac taaccataca tatttaaatt
      841 ttcaggtgct gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg agttaaaagg
      901 tattgatttt aaagaagatg gaaacattct tggacacaaa ttggaataca actataactc
      961 acacaatgta tacatcatgg cagacaaaca aaagaatgga atcaaagttg taagtttaaa
     1021 catgatttta ctaactaact aatctgattt aaattttcag aacttcaaaa ttagacacaa
     1081 cattgaagat ggaagcgttc aactagcaga ccattatcaa caaaatactc caattggcga
     1141 tggccctgtc cttttaccag acaaccatta cctgtcctac caatctgccc tttcgaaaga
     1201 tcccaacgaa aagagagacc acatggtcct tcttgagttt gtaacagctg ctgggattac
     1261 acatggcatg gatgaactat acaaatagca ttcgtagaat tgtaagttta aactttctca
     1321 tactaactaa cgcgctctat ttaaattttc agaattccaa ctgagcgccg gtcgctacca
     1381 ttaccaactt gtctggtgtc aaaaataata ggggccgctg tcatcagagt aagtttaaac
     1441 tgagttctac taactaacga gtaatattta aattttcagc atctcgcgcc cgtgcctctg
     1501 acttctaagt ccaattactc ttcaacatcc ctacatgctc tttctccctg tgctcccacc
     1561 ccctattttt gttattatca aaaaacttct cttaatttct ttgtttttta gcttctttta
     1621 agtcacctct aacaatgaaa ttgtgtagat tcaaaaatag aattaattcg taataaaaag
     1681 tcgaaaaaaa ttgtgctccc tccccccatt aataataatt ctatcccaaa atctacacaa
     1741 tgttctgtgt acacttctta tgttttttac ttctgataaa tttttttgaa acatcataga
     1801 aaaaaccgca cacaaaatac cttatcatat gttacgtttc agtttatgac cgcaattttt
     1861 atttcttcgc acgtctgggc ctctcatgac gtcaaatcat gctcatcgtg aaaaagtttt
     1921 ggagtatttt tggaattttt caatcaagtg aaagtttatg aaattaattt tcctgctttt
     1981 gctttttggg gtttccccta ttgtttgtca agatttcgag gacggcgttt ttcttgctaa
     2041 aatcacaagt attgatgagc acgatgcaag aaagatcgga agaaggtttg ggtttgaggc
     2101 tcagtggaag gtgagtagaa gttgataatt tgaaagtgga gtagtgtcta tggggttttt
     2161 gccttaaatg acagaataca ttcccaatat accaaacata actgtttcct actagtcggc
     2221 cgtacgggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     2281 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     2341 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     2401 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     2461 aataccgcat caggcggcct taagggcctc gtgatacgcc tatttttata ggttaatgtc
     2521 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     2581 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     2641 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     2701 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     2761 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     2821 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     2881 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     2941 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     3001 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     3061 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     3121 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     3181 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     3241 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     3301 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3361 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     3421 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3481 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     3541 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     3601 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3661 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3721 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3781 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3841 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3901 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3961 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4021 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4081 agatacctac agcgtgagca ttgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4141 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4201 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4261 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4321 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4381 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     4441 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     4501 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     4561 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     4621 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     4681 acaggaaaca gct
//
