LOCUS       Exported                4131 bp ds-DNA     circular SYN 21-MAR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4131)
  AUTHORS   NIH-NCI
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Mar 25, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4131
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(259..363)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(389..844)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        922..1899
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1934..1952
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      1984..1993
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1993..2016
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     2012..2069
                     /label=A692-pCS2+ empty-fragment
                     /note="Used for HIFI assembly and make this empty plasmid 
                     after digesting the pCS2+ hSRSF1 plasmid given by Wei Wu 
                     with EcoRI and XhoI"
     polyA_signal    2082..2216
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(2329..2347)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2368..2384)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2392..2408
                     /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)."
     promoter        complement(2416..2446)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2461..2482
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2770..3358)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(3529..4131,1..258))
                     /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"
ORIGIN
        1 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
       61 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
      121 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
      181 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
      241 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
      301 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
      361 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
      421 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
      481 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
      541 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
      601 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
      661 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
      721 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
      781 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
      841 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
      901 tcgctattac gccagtcgac cgccaattca atatggcgta tatggactca tgccaattca
      961 atatggtgga tctggacctg tgccaattca atatggcgta tatggactcg tgccaattca
     1021 atatggtgga tctggacccc agccaattca atatggcgga cttggcacca tgccaattca
     1081 atatggcgga cctggcactg tgccaactgg ggaggggtct acttggcacg gtgccaagtt
     1141 tgaggagggg tcttggccct gtgccaagtc cgccatattg aattggcatg gtgccaataa
     1201 tggcggccat attggctata tgccaggatc aatatatagg caatatccaa tatggcccta
     1261 tgccaatatg gctattggcc aggttcaata ctatgtattg gccctatgcc atatagtatt
     1321 ccatatatgg gttttcctat tgacgtagat agcccctccc aatgggcggt cccatatacc
     1381 atatatgggg cttcctaata ccgcccatag ccactccccc attgacgtca atggtctcta
     1441 tatatggtct ttcctattga cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc
     1501 ccattgacgt caattacggt aaatggcccg cctggctcaa tgcccattga cgtcaatagg
     1561 accacccacc attgacgtca atgggatggc tcattgccca ttcatatccg ttctcacgcc
     1621 ccctattgac gtcaatgacg gtaaatggcc cacttggcag tacatcaata tctattaata
     1681 gtaacttggc aagtacatta ctattggaag tacgccaggg tacattggca gtactcccat
     1741 tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa
     1801 tggggagggg caatgacgca aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc
     1861 taatgggagg tctatataag caatgctcgt ttagggaacc gccattctgc ctggggacgt
     1921 cggagcaagc ttgatttagg tgacactata gaatacaagc tacttgttct ttttgcagga
     1981 tcagccacca tggattacaa ggacgacgac gataagggat cccatcgatt cgaattcaag
     2041 gcctctcgag cctctagaac tatagtgagt cgtattacgt agatccagac atgataagat
     2101 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     2161 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     2221 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaat
     2281 tcgcggccgc ggcgccaatg cattgggccc ggtacccagc ttttgttccc tttagtgagg
     2341 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     2401 gctcacaatt ccacacaaca tacgagccgg gagcataaag tgtaaagcct ggggtgccta
     2461 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2521 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2581 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2641 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2701 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2761 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2821 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2881 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2941 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3001 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3061 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3121 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3181 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3241 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3301 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3361 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3421 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3481 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3541 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3601 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3661 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3721 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3781 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3841 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     3901 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     3961 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4021 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4081 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct c
//
