LOCUS       sequence-239-exp        4630 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 4630)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 4630)
  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: L4051
FEATURES             Location/Qualifiers
     source          1..4630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             527..547
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             569..589
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             611..631
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             653..673
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(716..744)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1492..1512
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2070..2092)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2192..2211
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2289..2307)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2375..2479
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2480..2548,2549..3340)
                     /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(2698..2717)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3511..4099
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4000..4019
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4253..4270
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4387..4408
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4423..4428,4429..4446,4447..4453)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4461..4477
                     /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     4466..4488
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4485..4501
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4485..4501
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggc cttggtcgac tctagaggat ccccggggat tggccaaagg
       61 acccaaaggt atgtttcgaa tgatactaac ataacataga acattttcag gaggaccctt
      121 gcttggctag caaaaatgca taaggttttg ctggcactgt tctttatctt tctggcacca
      181 gcatccgcac tggcagtctc cgaaccggcc tgcaggatcg attttttgca aattacgagc
      241 gttgtagggg gcggagcgat aggtcctata ggttttggta tatcatcatt cattcattca
      301 ttggtacatt catttaccca ccttcctctt tctgagcttc tctggagttc tgtgcttcct
      361 ttttccctta tctttatact gtaattttta actttcaggc attgattgga tccccgggat
      421 tggccaaagg acccaaaggt atgtttcgaa tgatactaac ataacataga acattttcag
      481 gaggaccctt gcttggaggg taccgagctc agaaaaaatg actgctccaa agaagaagcg
      541 taaggtaccg gcagaaaaaa tgactgctcc aaagaagaag cgtaaggtac cggcagaaaa
      601 aatgactgct ccaaagaaga agcgtaaggt accggcagaa aaaatgactg ctccaaagaa
      661 gaagcgtaag gtaccggtag aaaaaatgag taaaggagaa gaacttttca ctggagttgt
      721 cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga
      781 gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa
      841 actacctgtt ccatgggtaa gtttaaacat atatatacta actaaccctg attatttaaa
      901 ttttcagcca acacttgtca ctactttctg ttatggtgtt caatgcttct cgagataccc
      961 agatcatatg aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga
     1021 aagaactata tttttcaaag atgacgggaa ctacaagaca cgtaagttta aacagttcgg
     1081 tactaactaa ccatacatat ttaaattttc aggtgctgaa gtcaagtttg aaggtgatac
     1141 ccttgttaat agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg
     1201 acacaaattg gaatacaact ataactcaca caatgtatac atcatggcag acaaacaaaa
     1261 gaatggaatc aaagttgtaa gtttaaacat gattttacta actaactaat ctgatttaaa
     1321 ttttcagaac ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca
     1381 ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct
     1441 gtccacacaa tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttct
     1501 tgagtttgta acagctgctg ggattacaca tggcatggac gaactataca aaagcccaca
     1561 agctttggaa ttcgctagcc ggccatacaa gtaatccgga tgatcgacgc caacgtcgtt
     1621 gaattttcaa attttaaata ctgaatattt gttttttttc ctattattta tttattctct
     1681 ttgtgttttt tttcttgctt tctaaaaaat taattcaatc caaatctaaa catttttttt
     1741 tctctttccg tctcccaatt cgtattccgc tcctctcatc tgaacacaat gtgcaagttt
     1801 atttatcttc tcgctttcat ttcattagga cgtgggggga attggtggaa gggggaaaca
     1861 cacaaaagga tgatggaaat gaaataagga cacacaatat gcaacaacat tcaattcaga
     1921 aatatggagg aaggtttaaa agaaaacata aaaatatata gaggaggaag gaaaactagt
     1981 aaaaaataag caaagaaatt aggcgaacga tgagaattgt cctcgcttgg gccctttcgt
     2041 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     2101 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     2161 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     2221 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgg
     2281 ccttaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     2341 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     2401 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2461 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     2521 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     2581 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     2641 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     2701 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     2761 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     2821 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     2881 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     2941 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3001 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3061 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3121 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3181 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3241 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3301 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3361 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3421 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3481 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3541 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3601 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     3661 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3721 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3781 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3841 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3901 gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3961 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4021 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4081 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4141 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     4201 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     4261 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     4321 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     4381 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     4441 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     4501 ccatgattac gccaagctgt aagtttaaac atgatcttac taactaacta ttctcattta
     4561 aattttcaga gcttaaaaat ggctgaaatc actcacaacg atggatacgc taacaacttg
     4621 gaaatgaaat
//
