LOCUS       sequence-238-exp        4410 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 4410)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 4410)
  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: L4049
FEATURES             Location/Qualifiers
     source          1..4410
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             307..327
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             349..369
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             391..411
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             433..453
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(496..524)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1272..1292
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1850..1872)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1972..1991
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2069..2087)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2155..2259
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2260..2328,2329..3120)
                     /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(2478..2497)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3291..3879
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3780..3799
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4033..4050
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4167..4188
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4203..4208,4209..4226,4227..4233)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4241..4257
                     /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     4246..4268
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4265..4281
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4265..4281
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggc cttggtcgac tctagacact tttcagctac ctagatacat
       61 ggatatcccc gcctcccaat ccacccaccc agggaaaaag aagggctcgc cgaaaatcaa
      121 agttatctcc aggctcgcgc atcccaccga gcggttgact tctctccacc acttttcatt
      181 ttaaccctcg gggtacggga ttggccaaag gacccaaagg tatgtttcga atgatactaa
      241 cataacatag aacattttca ggaggaccct tcttggaggg taccgagctc agaaaaaatg
      301 actgctccaa agaagaagcg taaggtaccg gcagaaaaaa tgactgctcc aaagaagaag
      361 cgtaaggtac cggcagaaaa aatgactgct ccaaagaaga agcgtaaggt accggcagaa
      421 aaaatgactg ctccaaagaa gaagcgtaag gtaccggtag aaaaaatgag taaaggagaa
      481 gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac
      541 aaattttctg tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa
      601 tttatttgca ctactggaaa actacctgtt ccatgggtaa gtttaaacat atatatacta
      661 actaaccctg attatttaaa ttttcagcca acacttgtca ctactttctg ttatggtgtt
      721 caatgcttct cgagataccc agatcatatg aaacggcatg actttttcaa gagtgccatg
      781 cccgaaggtt atgtacagga aagaactata tttttcaaag atgacgggaa ctacaagaca
      841 cgtaagttta aacagttcgg tactaactaa ccatacatat ttaaattttc aggtgctgaa
      901 gtcaagtttg aaggtgatac ccttgttaat agaatcgagt taaaaggtat tgattttaaa
      961 gaagatggaa acattcttgg acacaaattg gaatacaact ataactcaca caatgtatac
     1021 atcatggcag acaaacaaaa gaatggaatc aaagttgtaa gtttaaacat gattttacta
     1081 actaactaat ctgatttaaa ttttcagaac ttcaaaatta gacacaacat tgaagatgga
     1141 agcgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt
     1201 ttaccagaca accattacct gtccacacaa tctgcccttt cgaaagatcc caacgaaaag
     1261 agagaccaca tggtccttct tgagtttgta acagctgctg ggattacaca tggcatggac
     1321 gaactataca aaagcccaca agctttggaa ttcgctagcc ggccatacaa gtaatccgga
     1381 tgatcgacgc caacgtcgtt gaattttcaa attttaaata ctgaatattt gttttttttc
     1441 ctattattta tttattctct ttgtgttttt tttcttgctt tctaaaaaat taattcaatc
     1501 caaatctaaa catttttttt tctctttccg tctcccaatt cgtattccgc tcctctcatc
     1561 tgaacacaat gtgcaagttt atttatcttc tcgctttcat ttcattagga cgtgggggga
     1621 attggtggaa gggggaaaca cacaaaagga tgatggaaat gaaataagga cacacaatat
     1681 gcaacaacat tcaattcaga aatatggagg aaggtttaaa agaaaacata aaaatatata
     1741 gaggaggaag gaaaactagt aaaaaataag caaagaaatt aggcgaacga tgagaattgt
     1801 cctcgcttgg gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     1861 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     1921 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     1981 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     2041 gaaaataccg catcaggcgg ccttaagggc ctcgtgatac gcctattttt ataggttaat
     2101 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     2161 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2221 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2281 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2341 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2401 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     2461 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     2521 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2581 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2641 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2701 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2761 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     2821 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2881 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2941 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3001 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3061 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3121 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     3181 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3241 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3301 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3361 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3421 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     3481 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3541 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3601 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3661 ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3721 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3781 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3841 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3901 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3961 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     4021 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     4081 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     4141 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     4201 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     4261 cacacaggaa acagctatga ccatgattac gccaagctgt aagtttaaac atgatcttac
     4321 taactaacta ttctcattta aattttcaga gcttaaaaat ggctgaaatc actcacaacg
     4381 atggatacgc taacaacttg gaaatgaaat
//
