LOCUS       Exported                4209 bp ds-DNA     circular SYN 14-JUN-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4209)
  AUTHORS   Dina
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Feb 17, 2022 from SnapGene Viewer 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4209
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    11..66
                     /label=RARE element
     promoter        83..279
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      130..265
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      296..305
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             302..1147
                     /codon_start=1
                     /product="EGFP destabilized by fusion to residues 422-461 
                     of mouse ornithine decarboxylase, giving an in vivo 
                     half-life of ~2 hours"
                     /label=d2EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKKLSHGFPPEVEEQDDGTLPMSCAQESGMDRHPAACASARINV
                     "
     polyA_site      1298..1353
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(1712..1728)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1736..1752
                     /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(1760..1790)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1805..1826
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1885..2080
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1931..2066
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2086..2220
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2459..3047)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3218..4078)
                     /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"
     promoter        complement(4079..4183)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacaattg cagttgggtc atttgaaggt tagcagcccg ggtagggttc accgaaagtt
       61 cactcgctcg agatctgcga tctgcatctc aattagtcag caaccatagt cccgccccta
      121 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatcgctga
      181 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
      241 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttggtac ctaaagccac
      301 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
      361 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
      421 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
      481 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
      541 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
      601 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
      661 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
      721 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
      781 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
      841 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
      901 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
      961 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagaa
     1021 gcttagccat ggcttcccgc cggaggtgga ggagcaggat gatggcacgc tgcccatgtc
     1081 ttgtgcccag gagagcggga tggaccgtca ccctgcagcc tgtgcttctg ctaggatcaa
     1141 tgtgtaggcg gccgcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg
     1201 gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg
     1261 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt
     1321 gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat
     1381 catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc catatgctgg
     1441 ctgccatgaa caaaggtggc tataaagagg tcatcagtat atgaaacagc cccctgctgt
     1501 ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt
     1561 tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac tagccagatt
     1621 tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat gaagatccct
     1681 cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     1741 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     1801 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     1861 gaaacctgtc gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct
     1921 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     1981 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     2041 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctaactt gtttattgca
     2101 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     2161 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     2221 cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2281 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2341 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2401 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2461 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2521 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2581 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2641 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2701 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2761 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2821 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2881 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2941 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3001 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3061 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3121 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3181 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3241 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3301 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3361 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3421 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3481 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3541 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3601 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3661 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3721 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3781 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3841 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3901 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3961 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4021 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4081 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4141 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4201 tgccacctg
//
