LOCUS       sequence-232-exp        4280 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 4280)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 4280)
  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: L4041
FEATURES             Location/Qualifiers
     source          1..4280
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             177..197
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             219..239
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             261..281
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             303..323
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(366..394)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1142..1162
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1720..1742)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1842..1861
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1939..1957)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2025..2129
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2130..2198,2199..2990)
                     /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(2348..2367)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3161..3749
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3650..3669
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3903..3920
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4037..4058
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4073..4078,4079..4096,4097..4103)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4111..4127
                     /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     4116..4138
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4135..4151
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4135..4151
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ccgcggtttt ggcgcggatc ctatcgattc
       61 gcggccgctg tacaccccca aaggacccaa aggtatgttt cgaatgatac taacataaca
      121 tagaacattt tcaggaggac ccttggaggg taccgagctc agaaaaaatg actgctccaa
      181 agaagaagcg taaggtaccg gcagaaaaaa tgactgctcc aaagaagaag cgtaaggtac
      241 cggcagaaaa aatgactgct ccaaagaaga agcgtaaggt accggcagaa aaaatgactg
      301 ctccaaagaa gaagcgtaag gtaccggtag aaaaaatgag taaaggagaa gaacttttca
      361 ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg
      421 tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca
      481 ctactggaaa actacctgtt ccatgggtaa gtttaaacat atatatacta actaaccctg
      541 attatttaaa ttttcagcca acacttgtca ctactttctg ttatggtgtt caatgcttct
      601 cgagataccc agatcatatg aaacggcatg actttttcaa gagtgccatg cccgaaggtt
      661 atgtacagga aagaactata tttttcaaag atgacgggaa ctacaagaca cgtaagttta
      721 aacagttcgg tactaactaa ccatacatat ttaaattttc aggtgctgaa gtcaagtttg
      781 aaggtgatac ccttgttaat agaatcgagt taaaaggtat tgattttaaa gaagatggaa
      841 acattcttgg acacaaattg gaatacaact ataactcaca caatgtatac atcatggcag
      901 acaaacaaaa gaatggaatc aaagttgtaa gtttaaacat gattttacta actaactaat
      961 ctgatttaaa ttttcagaac ttcaaaatta gacacaacat tgaagatgga agcgttcaac
     1021 tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca
     1081 accattacct gtccacacaa tctgcccttt cgaaagatcc caacgaaaag agagaccaca
     1141 tggtccttct tgagtttgta acagctgctg ggattacaca tggcatggac gaactataca
     1201 aaagcccaca agctttggaa ttcgctagcc ggccatacaa gtaatccgga tgatcgacgc
     1261 caacgtcgtt gaattttcaa attttaaata ctgaatattt gttttttttc ctattattta
     1321 tttattctct ttgtgttttt tttcttgctt tctaaaaaat taattcaatc caaatctaaa
     1381 catttttttt tctctttccg tctcccaatt cgtattccgc tcctctcatc tgaacacaat
     1441 gtgcaagttt atttatcttc tcgctttcat ttcattagga cgtgggggga attggtggaa
     1501 gggggaaaca cacaaaagga tgatggaaat gaaataagga cacacaatat gcaacaacat
     1561 tcaattcaga aatatggagg aaggtttaaa agaaaacata aaaatatata gaggaggaag
     1621 gaaaactagt aaaaaataag caaagaaatt aggcgaacga tgagaattgt cctcgcttgg
     1681 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     1741 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     1801 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     1861 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     1921 catcaggcgg ccttaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     1981 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     2041 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2101 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     2161 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     2221 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     2281 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     2341 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     2401 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     2461 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     2521 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     2581 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     2641 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     2701 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     2761 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     2821 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     2881 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     2941 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     3001 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     3061 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     3121 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3181 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3241 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3301 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3361 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3421 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3481 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3541 tacagcgtga gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3601 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3661 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3721 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3781 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3841 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     3901 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     3961 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     4021 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     4081 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     4141 acagctatga ccatgattac gccaagctgt aagtttaaac atgatcttac taactaacta
     4201 ttctcattta aattttcaga gcttaaaaat ggctgaaatc actcacaacg atggatacgc
     4261 taacaacttg gaaatgaaat
//
