LOCUS       sequence-237-exp        4247 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 4247)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 4247)
  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: L4048
FEATURES             Location/Qualifiers
     source          1..4247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             144..164
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             186..206
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             228..248
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             270..290
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(333..361)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1109..1129
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1687..1709)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1809..1828
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1906..1924)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1992..2096
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2097..2165,2166..2957)
                     /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(2315..2334)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3128..3716
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3617..3636
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3870..3887
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4004..4025
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4040..4045,4046..4063,4064..4070)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4078..4094
                     /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     4083..4105
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4102..4118
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4102..4118
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactagagg atccccggga ttggccaaag gacccaaagg
       61 tatgtttcga atgatactaa cataacatag aacattttca ggaggaccct tggagggtac
      121 cgagctcaga aaaaatgact gctccaaaga agaagcgtaa ggtaccggca gaaaaaatga
      181 ctgctccaaa gaagaagcgt aaggtaccgg cagaaaaaat gactgctcca aagaagaagc
      241 gtaaggtacc ggcagaaaaa atgactgctc caaagaagaa gcgtaaggta ccggtagaaa
      301 aaatgagtaa aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg
      361 gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg
      421 gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcca tgggtaagtt
      481 taaacatata tatactaact aaccctgatt atttaaattt tcagccaaca cttgtcacta
      541 ctttctgtta tggtgttcaa tgcttctcga gatacccaga tcatatgaaa cggcatgact
      601 ttttcaagag tgccatgccc gaaggttatg tacaggaaag aactatattt ttcaaagatg
      661 acgggaacta caagacacgt aagtttaaac agttcggtac taactaacca tacatattta
      721 aattttcagg tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa
      781 aaggtattga ttttaaagaa gatggaaaca ttcttggaca caaattggaa tacaactata
      841 actcacacaa tgtatacatc atggcagaca aacaaaagaa tggaatcaaa gttgtaagtt
      901 taaacatgat tttactaact aactaatctg atttaaattt tcagaacttc aaaattagac
      961 acaacattga agatggaagc gttcaactag cagaccatta tcaacaaaat actccaattg
     1021 gcgatggccc tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcga
     1081 aagatcccaa cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga
     1141 ttacacatgg catggacgaa ctatacaaaa gcccacaagc tttggaattc gctagccggc
     1201 catacaagta atccggatga tcgacgccaa cgtcgttgaa ttttcaaatt ttaaatactg
     1261 aatatttgtt ttttttccta ttatttattt attctctttg tgtttttttt cttgctttct
     1321 aaaaaattaa ttcaatccaa atctaaacat ttttttttct ctttccgtct cccaattcgt
     1381 attccgctcc tctcatctga acacaatgtg caagtttatt tatcttctcg ctttcatttc
     1441 attaggacgt ggggggaatt ggtggaaggg ggaaacacac aaaaggatga tggaaatgaa
     1501 ataaggacac acaatatgca acaacattca attcagaaat atggaggaag gtttaaaaga
     1561 aaacataaaa atatatagag gaggaaggaa aactagtaaa aaataagcaa agaaattagg
     1621 cgaacgatga gaattgtcct cgcttgggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     1681 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     1741 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     1801 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     1861 gcacagatgc gtaaggagaa aataccgcat caggcggcct taagggcctc gtgatacgcc
     1921 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     1981 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2041 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     2101 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2161 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     2221 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     2281 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     2341 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     2401 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     2461 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     2521 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     2581 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     2641 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     2701 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     2761 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     2821 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     2881 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     2941 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     3001 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3061 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3121 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3181 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3241 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     3301 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3361 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3421 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3481 gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc gccacgcttc
     3541 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3601 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3661 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     3721 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     3781 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     3841 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     3901 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     3961 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     4021 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     4081 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagctgtaag
     4141 tttaaacatg atcttactaa ctaactattc tcatttaaat tttcagagct taaaaatggc
     4201 tgaaatcact cacaacgatg gatacgctaa caacttggaa atgaaat
//
