LOCUS       sequence-235-exp        4248 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 4248)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 4248)
  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: L4046
FEATURES             Location/Qualifiers
     source          1..4248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             145..165
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             187..207
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             229..249
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             271..291
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(334..362)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1110..1130
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1688..1710)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1810..1829
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1907..1925)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1993..2097
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2098..2166,2167..2958)
                     /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(2316..2335)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3129..3717
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3618..3637
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3871..3888
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4005..4026
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4041..4046,4047..4064,4065..4071)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4079..4095
                     /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     4084..4106
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4103..4119
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4103..4119
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ggatccccgg gattggccaa aggacccaaa
       61 ggtatgtttc gaatgatact aacataacat agaacatttt caggaggacc cttgagggta
      121 ccgagctcag aaaaaatgac tgctccaaag aagaagcgta aggtaccggc agaaaaaatg
      181 actgctccaa agaagaagcg taaggtaccg gcagaaaaaa tgactgctcc aaagaagaag
      241 cgtaaggtac cggcagaaaa aatgactgct ccaaagaaga agcgtaaggt accggtagaa
      301 aaaatgagta aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat
      361 ggtgatgtta atgggcacaa attttctgtc agtggagagg gtgaaggtga tgcaacatac
      421 ggaaaactta cccttaaatt tatttgcact actggaaaac tacctgttcc atgggtaagt
      481 ttaaacatat atatactaac taaccctgat tatttaaatt ttcagccaac acttgtcact
      541 actttctgtt atggtgttca atgcttctcg agatacccag atcatatgaa acggcatgac
      601 tttttcaaga gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat
      661 gacgggaact acaagacacg taagtttaaa cagttcggta ctaactaacc atacatattt
      721 aaattttcag gtgctgaagt caagtttgaa ggtgataccc ttgttaatag aatcgagtta
      781 aaaggtattg attttaaaga agatggaaac attcttggac acaaattgga atacaactat
      841 aactcacaca atgtatacat catggcagac aaacaaaaga atggaatcaa agttgtaagt
      901 ttaaacatga ttttactaac taactaatct gatttaaatt ttcagaactt caaaattaga
      961 cacaacattg aagatggaag cgttcaacta gcagaccatt atcaacaaaa tactccaatt
     1021 ggcgatggcc ctgtcctttt accagacaac cattacctgt ccacacaatc tgccctttcg
     1081 aaagatccca acgaaaagag agaccacatg gtccttcttg agtttgtaac agctgctggg
     1141 attacacatg gcatggacga actatacaaa agcccacaag ctttggaatt cgctagccgg
     1201 ccatacaagt aatccggatg atcgacgcca acgtcgttga attttcaaat tttaaatact
     1261 gaatatttgt tttttttcct attatttatt tattctcttt gtgttttttt tcttgctttc
     1321 taaaaaatta attcaatcca aatctaaaca tttttttttc tctttccgtc tcccaattcg
     1381 tattccgctc ctctcatctg aacacaatgt gcaagtttat ttatcttctc gctttcattt
     1441 cattaggacg tggggggaat tggtggaagg gggaaacaca caaaaggatg atggaaatga
     1501 aataaggaca cacaatatgc aacaacattc aattcagaaa tatggaggaa ggtttaaaag
     1561 aaaacataaa aatatataga ggaggaagga aaactagtaa aaaataagca aagaaattag
     1621 gcgaacgatg agaattgtcc tcgcttgggc cctttcgtct cgcgcgtttc ggtgatgacg
     1681 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     1741 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     1801 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     1861 cgcacagatg cgtaaggaga aaataccgca tcaggcggcc ttaagggcct cgtgatacgc
     1921 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     1981 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2041 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     2101 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     2161 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     2221 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     2281 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     2341 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     2401 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     2461 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     2521 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     2581 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     2641 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     2701 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     2761 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     2821 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     2881 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     2941 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     3001 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     3061 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3121 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3181 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3241 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     3301 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3361 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3421 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3481 cgaacgacct acaccgaact gagataccta cagcgtgagc attgagaaag cgccacgctt
     3541 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3601 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3661 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     3721 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     3781 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     3841 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     3901 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     3961 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     4021 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     4081 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctgtaa
     4141 gtttaaacat gatcttacta actaactatt ctcatttaaa ttttcagagc ttaaaaatgg
     4201 ctgaaatcac tcacaacgat ggatacgcta acaacttgga aatgaaat
//
