LOCUS       Exported                7570 bp ds-DNA     circular SYN 10-SEP-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7570)
  AUTHORS   Enzo Mameli
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Sep 10, 2021 from SnapGene 5.1.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7570
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             201..1061
                     /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"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the 
                     presenceof cAMP."
     promoter        2144..2174
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator 
                     toinhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2276..2309
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Grothet
                     al., 2000)"
     promoter        2319..2462
                     /label=U6 (AGAP013695)
                     /label=U6 (AGAP013695 L)
                     /note="polIII promoter from AGAP013695 Konin et al. (long 
                     form) tested in An. gambiae cells. Tested in adult 
                     mosquitos in Hammond et al."
     misc_RNA        2481..2556
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2565..2599
                     /label=3'Flank AGAP0013695
     misc_feature    2600..2786
                     /label=3'end snRNAU696AB Dmel
     promoter        2845..5354
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             5388..6104
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKGD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             6108..6161
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus 
                     capsidprotein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             6186..6785
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    6812..6933
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    7127..7160
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Grothet
                     al., 2000)"
     primer_bind     complement(7176..7192)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt gcatgcctgc aggtcgactc tagaggatcc cccgggtgcc
     2281 agggcgtgcc cttgggctcc ccgggcgcgt acttcgaatt tgtatgcgtg cgcttgaagg
     2341 gttgatcgga accttacaac agttgtagct atacggctgc gtgtggcttc taacgttatc
     2401 catcgctaga agtgaaacga atgtgcgtag gtatatatat gaaatggagt tgctctctgc
     2461 ttgggtcttc gagaagacct gttttagagc tagaaatagc aagttaaaat aaggctagtc
     2521 cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttttgacact tttccattcc
     2581 tcctgcttca tttgacgact tttttgccta cctggagcct gagagttgtt caataaaata
     2641 aaaatgtttc gtttttttgc tttcgccagt atttattatt tttcatcaat atgtattcaa
     2701 tttggtatgt atttagtaat tgtaatatat agacaatggt tttccgttga cgtacataca
     2761 tctgacgtgt gtttatttag acataaggta cccagcatgc aattctatat tctaaaaaca
     2821 caaatgatac ttctaaaaaa aaatcatgaa tggcatcaac tctgaatcaa atctttgcag
     2881 atgcacctac ttctcatttc cactgtcaca tcatttttcc agatctcgct gcctgttatg
     2941 tggcccacaa accaagacac gttttatggc cattaaagct ggctgatcgt cgccaaacac
     3001 caaatacata atgaatatgt acacattcga gaaagaagcg atcaaagaag catcttcggg
     3061 cggagtagga gaatgcggag gagaaggaga acgagctgat ctagtatctc tccacaatcc
     3121 aatgccaact gaccaactgg ccatattcgg agcaatttga agccaatttc catcgcctgg
     3181 cgatcgctcc attcttggct atatgttttt caccgttacc cggggccatt ttcaaagact
     3241 cgtcggcaag ataagattgt gtcactcgct gtctctcttc atttgtcgaa gaatgctgag
     3301 gaatttcgcg atgacgtcgg cgagtatttt gaagaatgag aataatttgt atttatacga
     3361 aaatcagtta gtggaatttt ctacaaaaac atgttatcta tagataattt tgttgcaaaa
     3421 tatgttgact atgacaaaga ttgtatgtat atacctttaa tgtattctca ttttcttatg
     3481 tatttataat ggcaatgatg atactgatga tattttaaga tgatgccaga ccaaaaggct
     3541 tgaatttctg cgtcttttgc cgaacgcagt gcatgtgcaa ttgttgtttt ttggaatatt
     3601 caattttcgg actgtccgct ttgatttcag tttcttggct tattcaaaaa gcaaagtaaa
     3661 gccaaaaaag cgagatggca ataccaaatg cggcaaaacg gtagtggaag gaaaggggtg
     3721 cggggcagcg gaaggaaggg tggggcgggg cgtggcgggg tctgtggctg ggcgcgacgt
     3781 caccgacgtt ggagccactc ctttgaccat gtgtgcgtgt gtgtattatt cgtgtctcgc
     3841 cactcgccgg ttgttttttt ctttttatgc tgcgctctct ctagcgccat ctcgcttacg
     3901 catgctcaac gcaccgcatg ttgccgtttc cttttatgcg tcattttggc tcgaaatagg
     3961 caattattta aacaaagatt agtcaacgaa aacgctaaaa taaataagtc tacaatatgg
     4021 ttacttattg ccatgtgtgt gcagccaacg atagcaacaa aagcaacaac acaggtggct
     4081 ttccctcttt cactttttgt ttgcaagccg cgtgcgagca agacggcacg accggcaaac
     4141 gcaattacgc tgacaaagag cagacgaagt tttggcgaaa aacatcaagg cgcctgatac
     4201 gaatgcattt gcaataacaa ttgcgatatt taatattgtt tatgaagctg tttgacttca
     4261 aaacacacaa aaaaaaaaat aaaacaaatt atttgaaaga gaattaggaa tcggacgctt
     4321 atcgttaggg taacaacaag aaatgcttac tgagtcacag cctctggaaa actgccgcaa
     4381 gccagagaga gagagaaaaa gagggagagc agcttagacc gcatgtgctt gtgtgtgagg
     4441 cgtctctctc ttcgtctctg ttgcgcaaac gcatagactg cactgagaaa atcgattacc
     4501 tattttttat gaatgaatat ttgcactatt actattcaaa actattaaga tagcaatcac
     4561 attcaatagc caaatactat accacctgag cgatgcaacg aaatgatcaa tttgagcaaa
     4621 aatgctgcat atttaggacg gcatcattat agaaatgctt cttgctgtgt acttttctct
     4681 cgtctggcag ctgtttcgcc gttattgtta aaaccggctt aagttaggtg tgttttctac
     4741 gactagtgaa tgccctacta gaagatgtgt gttgcacaaa atgtccctgg aataaccaat
     4801 ttgaagtgca gatagcagta aacgtaagct aatatgaata ttatttaact gtaatgtttt
     4861 aatatcgctg gacattacta ataaacccac tataaacaca tgtacatatg tatgttttgg
     4921 catacaatga gtagttgggg aaaaaatgtg taaaagcacc gtgaccatca cagcataaag
     4981 ataaccagct gaagtatcga atatgagtaa cccccaaatt gaatcacatg ccgcaactga
     5041 taggacccat ggaagtacac tcttcatggc gatatacaag acacacacaa gcacgaacac
     5101 ccagttgcgg aggaaattct ccgtaaatga aaacccaatc ggcgaacaat tcatacccat
     5161 atatggtaaa agttttgaac gcgacttgag agcggagagc attgcggctg ataaggtttt
     5221 agcgctaagc gggctttata aaacgggctg cgggaccagt tttcatatca ctaccgtttg
     5281 agttcttgtg ctgtgtggat actcctcccg acacaaagcc gctccatcag ccagcagtcg
     5341 tctaatccag agaccccgga tccagatgaa ttcgcccttg tttaaacatg gtgagcaagg
     5401 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     5461 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     5521 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     5581 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     5641 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aagggcgacg
     5701 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     5761 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     5821 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     5881 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     5941 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     6001 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     6061 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagcttgag ggcagaggaa
     6121 gccttctaac atgcggtgac gtggaggaga atcccggccc tgctagcggt agcggcagcg
     6181 gtagcatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg
     6241 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc
     6301 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     6361 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     6421 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     6481 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     6541 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     6601 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     6661 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     6721 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     6781 cctgactcga gggatccgaa ttcagacatg ataagataca ttgatgagtt tggacaaacc
     6841 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     6901 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     6961 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     7021 ggtatggctg attatgatca gtcgaccgat gcccttgaga gccttcaacc cagtcagctc
     7081 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atggtacgcg cccggggagc
     7141 ccaagggcac gccctggcac ccgaagggcg aattcactgg ccgtcgtttt acaacgtcgt
     7201 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     7261 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     7321 aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     7381 cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     7441 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     7501 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     7561 aaacgcgcga
//
