LOCUS       sequence-233-exp        4674 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 4674)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 4674)
  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: L4042
FEATURES             Location/Qualifiers
     source          1..4674
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             571..591
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             613..633
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             655..675
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             697..717
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(760..788)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1536..1556
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2114..2136)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2236..2255
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2333..2351)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2419..2523
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2524..2592,2593..3384)
                     /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(2742..2761)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3555..4143
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4044..4063
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4297..4314
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4431..4452
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4467..4472,4473..4490,4491..4497)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4505..4521
                     /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     4510..4532
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4529..4545
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4529..4545
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ggatcaccaa aaacggaacg ttgagctgga
       61 cggaaatagt ggtaaagtga catgattata gtttgaagat ttctaatttc acaattagag
      121 caaatgttgt tcggtattta ttttcaacgg tatttatact attttccacc tttttctaga
      181 acattcgagc tgcttgttgc aaaaggaggg cgactcacat tcggtacatg gaaaagtagt
      241 gtacacaata aagagaccca gatacatttt ccgtctgcgt ctctttgcac ccaccgggag
      301 tattttcaaa cgaatgcatc taggaccttc tagaacattc tgtaaggctg cagaatgcgg
      361 gtatataagg aaagcgggct cagaggaagc caacacgctt tgttctagtg catctaaaaa
      421 acttcgaaaa tcctcatcgg gatcctatcg attcgcggcc gctgtacacc cccaaaggac
      481 ccaaaggtat gtttcgaatg atactaacat aacatagaac attttcagga ggacccttgg
      541 agggtaccga gctcagaaaa aatgactgct ccaaagaaga agcgtaaggt accggcagaa
      601 aaaatgactg ctccaaagaa gaagcgtaag gtaccggcag aaaaaatgac tgctccaaag
      661 aagaagcgta aggtaccggc agaaaaaatg actgctccaa agaagaagcg taaggtaccg
      721 gtagaaaaaa tgagtaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa
      781 ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca
      841 acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg
      901 gtaagtttaa acatatatat actaactaac cctgattatt taaattttca gccaacactt
      961 gtcactactt tctgttatgg tgttcaatgc ttctcgagat acccagatca tatgaaacgg
     1021 catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac tatatttttc
     1081 aaagatgacg ggaactacaa gacacgtaag tttaaacagt tcggtactaa ctaaccatac
     1141 atatttaaat tttcaggtgc tgaagtcaag tttgaaggtg atacccttgt taatagaatc
     1201 gagttaaaag gtattgattt taaagaagat ggaaacattc ttggacacaa attggaatac
     1261 aactataact cacacaatgt atacatcatg gcagacaaac aaaagaatgg aatcaaagtt
     1321 gtaagtttaa acatgatttt actaactaac taatctgatt taaattttca gaacttcaaa
     1381 attagacaca acattgaaga tggaagcgtt caactagcag accattatca acaaaatact
     1441 ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc
     1501 ctttcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct
     1561 gctgggatta cacatggcat ggacgaacta tacaaaagcc cacaagcttt ggaattcgct
     1621 agccggccat acaagtaatc cggatgatcg acgccaacgt cgttgaattt tcaaatttta
     1681 aatactgaat atttgttttt tttcctatta tttatttatt ctctttgtgt tttttttctt
     1741 gctttctaaa aaattaattc aatccaaatc taaacatttt tttttctctt tccgtctccc
     1801 aattcgtatt ccgctcctct catctgaaca caatgtgcaa gtttatttat cttctcgctt
     1861 tcatttcatt aggacgtggg gggaattggt ggaaggggga aacacacaaa aggatgatgg
     1921 aaatgaaata aggacacaca atatgcaaca acattcaatt cagaaatatg gaggaaggtt
     1981 taaaagaaaa cataaaaata tatagaggag gaaggaaaac tagtaaaaaa taagcaaaga
     2041 aattaggcga acgatgagaa ttgtcctcgc ttgggccctt tcgtctcgcg cgtttcggtg
     2101 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     2161 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     2221 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     2281 aaataccgca cagatgcgta aggagaaaat accgcatcag gcggccttaa gggcctcgtg
     2341 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     2401 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     2461 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     2521 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     2581 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     2641 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     2701 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     2761 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     2821 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     2881 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     2941 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     3001 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     3061 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     3121 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     3181 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     3241 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     3301 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3361 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     3421 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     3481 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3541 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3601 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3661 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     3721 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3781 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3841 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3901 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagcattg agaaagcgcc
     3961 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4021 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     4081 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4141 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     4201 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     4261 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     4321 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     4381 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     4441 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     4501 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     4561 ctgtaagttt aaacatgatc ttactaacta actattctca tttaaatttt cagagcttaa
     4621 aaatggctga aatcactcac aacgatggat acgctaacaa cttggaaatg aaat
//
