LOCUS       sequence-129-exp        4835 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 4835)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1997
REFERENCE   2  (bases 1 to 4835)
  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: L2684
FEATURES             Location/Qualifiers
     source          1..4835
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             583..603
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(646..674)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1422..1442
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2409..2431)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2531..2550
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2628..2646)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2714..2818
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2819..2887,2888..3679)
                     /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(3037..3056)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3850..4438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4339..4358
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4592..4609
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4726..4747
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4762..4767,4768..4785,4786..4792)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4800..4816
                     /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     4805..4827
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(4824..4835,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(4824..4835,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tcaccaaaaa
       61 cggaacgttg agctggacgg aaatagtggt aaagtgacat gattatagtt tgaagatttc
      121 taatttcaca attagagcaa atgttgttcg gtatttattt tcaacggtat ttatactatt
      181 ttccaccttt ttctagaaca ttcgagctgc ttgttgcaaa aggagggcga ctcacattcg
      241 gtacatggaa aagtagtgta cacaataaag agacccagat acattttccg tctgcgtctc
      301 tttgcaccca ccgggagtat tttcaaacga atgcatctag gaccttctag aacattctgt
      361 aaggctgcag aatgcgggta tataaggaaa gcgggctcag aggaagccaa cacgctttgt
      421 tctagtgcat ctaaaaaact tcgaaaatcc tcatcggatc cccgggattg gccaaaggac
      481 ccaaaggtat gtttcgaatg atactaacat aacatagaac attttcagga ggacccttgg
      541 ctagaggatc cccgggtacc gagctcagaa aaaatgactg ctccaaagaa gaagcgtaag
      601 gtaccggtag aaaaaatgag taaaggagaa gaacttttca ctggagttgt cccaattctt
      661 gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga gggtgaaggt
      721 gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa actacctgtt
      781 ccatgggtaa gtttaaacat atatatacta actaaccctg attatttaaa ttttcagcca
      841 acacttgtca ctactttctg ttatggtgtt caatgcttct cgagataccc agatcatatg
      901 aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga aagaactata
      961 tttttcaaag atgacgggaa ctacaagaca cgtaagttta aacagttcgg tactaactaa
     1021 ccatacatat ttaaattttc aggtgctgaa gtcaagtttg aaggtgatac ccttgttaat
     1081 agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg acacaaattg
     1141 gaatacaact ataactcaca caatgtatac atcatggcag acaaacaaaa gaatggaatc
     1201 aaagttgtaa gtttaaacat gattttacta actaactaat ctgatttaaa ttttcagaac
     1261 ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa
     1321 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa
     1381 tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttct tgagtttgta
     1441 acagctgctg ggattacaca tggcatggat gaactataca aatagcattc gtagaattcc
     1501 aactgagcgc cggtcgctac cattaccaac ttgtctggtg tcaaaaataa taggggccgc
     1561 tgtcatcaga gtaagtttaa actgagttct actaactaac gagtaatatt taaattttca
     1621 gcatctcgcg cccgtgcctc tgacttctaa gtccaattac tcttcaacat ccctacatgc
     1681 tctttctccc tgtgctccca ccccctattt ttgttattat caaaaaactt ctcttaattt
     1741 ctttgttttt tagcttcttt taagtcacct ctaacaatga aattgtgtag attcaaaaat
     1801 agaattaatt cgtaataaaa agtcgaaaaa aattgtgctc cctcccccca ttaataataa
     1861 ttctatccca aaatctacac aatgttctgt gtacacttct tatgtttttt acttctgata
     1921 aatttttttg aaacatcata gaaaaaaccg cacacaaaat accttatcat atgttacgtt
     1981 tcagtttatg accgcaattt ttatttcttc gcacgtctgg gcctctcatg acgtcaaatc
     2041 atgctcatcg tgaaaaagtt ttggagtatt tttggaattt ttcaatcaag tgaaagttta
     2101 tgaaattaat tttcctgctt ttgctttttg gggtttcccc tattgtttgt caagatttcg
     2161 aggacggcgt ttttcttgct aaaatcacaa gtattgatga gcacgatgca agaaagatcg
     2221 gaagaaggtt tgggtttgag gctcagtgga aggtgagtag aagttgataa tttgaaagtg
     2281 gagtagtgtc tatggggttt ttgccttaaa tgacagaata cattcccaat ataccaaaca
     2341 taactgtttc ctactagtcg gccgtacggg ccctttcgtc tcgcgcgttt cggtgatgac
     2401 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     2461 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     2521 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     2581 ccgcacagat gcgtaaggag aaaataccgc atcaggcggc cttaagggcc tcgtgatacg
     2641 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     2701 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     2761 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     2821 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     2881 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     2941 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3001 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     3061 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     3121 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     3181 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     3241 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     3301 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     3361 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     3421 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     3481 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     3541 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     3601 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     3661 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     3721 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     3781 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3841 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3901 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3961 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     4021 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4081 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4141 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4201 gcgaacgacc tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct
     4261 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4321 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4381 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4441 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     4501 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     4561 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     4621 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     4681 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     4741 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     4801 tgtgagcgga taacaatttc acacaggaaa cagct
//
