LOCUS       Exported                8221 bp ds-DNA     circular SYN 21-MAY-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8221)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jan 20, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8221
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    54..2717
                     /label=tag-168 promoter
     primer_bind     2796..2816
                     /label=pSF11-Fw-Seq
     primer_bind     2808..2857
                     /label=0507-619-F
     misc_feature    2834..2837
                     /label=optimized sequence for ribosomal binding site
     misc_feature    2834..2837
                     /label=codon adapted insert for smGFP-myc
                     /note="codon adapted by 
                     https://worm.mpi-cbg.de/codons/cgi-bin/optimize.py
                     Intron * 2 (3 requested, but 1 is inside myc regions and 
                     thus removed)
                     CAI set to 0.60
                     Check all 3 boxes
                     "
     CDS             2838..3869
                     /codon_start=1
                     /label=wemiRFP670
                     /translation="MAEGSVARQPDLLTCEHEEIHLAGSIQPHGALLVVSEHDHRVIQA
                     SANAAEFLNLGSVLGVPLAEIDGDLLIKVSLNIYILTNPDYLNFQILPHLDPTAEGMPV
                     AVRCRIGNPSTEYCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALC
                     DDTVLLFQQCTGYDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSTVPQMARQLY
                     VRQRVRVLVDVTYQPVPLEPRLSPLTGRDLDMSGCFLRSMSPCHLQFLKVSLNSSVLTN
                     HTYLNFQDMGVRATLAVSLVVGGKLWGLVVCHHYLPRFIRFELRAICKRLAERIATRIT
                     ALES"
     misc_feature    3057..3107
                     /label=intron-1
     misc_feature    3651..3701
                     /label=intron-2
     primer_bind     complement(3849..3890)
                     /label=0507-619-R
     CDS             3870..3890
                     /codon_start=1
                     /label=Linker
                     /translation="GYDIGSG"
     primer_bind     3870..3890
                     /label=0607-wHO1-F
     CDS             3891..3944
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3945..4964
                     /codon_start=1
                     /label=HO1
                     /translation="MERPQPDSMPQDLSEALKEATKEVHTQAENAEFMRNFQKGQVTRD
                     GFKLVMASLYHIYVALEEEIERNKVSLNIYILTNPDYLNFQESPVFAPVYFPEELHRKA
                     ALEQDLAFWYGPRWQEVIPYTPAMQRYVKRLHEVGRTEPELLVAHAYTRYLGDLSGGQV
                     LKVSLNSSVLTNHTYLNFQKIAQKALDLPSSGEGLAFFTFPNIASATKFKQLYRSRMNS
                     LEMTPAVRQRVIEEAKVSLNMILLTN*SDLNFQTAFLLNIQLFEELQELLTHDTKDQSP
                     SRAPGLRQRASNKVQDSAPVETPRGKPPLNTRSQAPLLRWVLTLSFLVATVAVGLYAM"
     misc_feature    4152..4202
                     /label=Intron-1
     primer_bind     complement(4217..4239)
                     /label=wHO1-R
     primer_bind     complement(4332..4353)
                     /label=wHO1-R-seq2
     misc_feature    4440..4490
                     /label=Intron-2
     misc_feature    4659..4709
                     /label=Intron-3
     primer_bind     complement(4941..4990)
                     /label=0507-wHO1-R
     primer_bind     complement(5006..5022)
                     /label=pSF11-Rv-Seq
     promoter        6100..6204
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6205..7065
                     /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"
     CDS             6403..7062
                     /label=AmpR
     rep_origin      7160..7842
                     /label=ColE1 origin
     rep_origin      7236..7824
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7921..7939)
                     /label=0507-619-R
     protein_bind    8112..8133
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8148..8178
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     misc_binding    8182..8204
                     /label=LacO
     protein_bind    8186..8202
                     /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     join(8210..8221,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcg cggccgcact gactgggccg gccggatcca
       61 taggtatgcc tgtcattctc tatttttaaa cgttcctgca gcagtagcac aacagagcca
      121 gcagttttcc tcttcttctc catcttcttc tgcttcttcc tctcttccca aaaaaaaaaa
      181 aaaaaaaaaa gtgagggggc tcactgcccc cccccccctc ccccccccct cttctccttg
      241 aagctcatcc agacgtcgct gtcctcctcc tcctcgtccc cggggcgctg tgtgtgtgtg
      301 tgtgtgctcc gtgagcatgt agcagcttca gcccatttaa aggcgcacac tctctgcatc
      361 tctgcgtgtc tcgacactcg ttttttctct tcttttctgc tgcacacaca cacacacaca
      421 cacacacgct catgccagag cacttcttgc tccggggggc tccgcgcgga gcccgagagg
      481 acctcctcct cctttttctt cttcgctttt tagttcttct tgcctcggtc tcgccccttg
      541 ttttgctctc catgtagtat atacatctgg gtgtgtgtgt gtgtgtgcac ctttggagca
      601 ttttttgaac ttttccagaa ctttggagca acattttcag cttgttcagt gcacttttta
      661 ggctgaaaat cctattttta ggttttcgaa agtttttttg gagcattttt cgtgcaattt
      721 aggtgcaaaa ttaaaaaaaa gggttttgta gtgaattaaa atttttttga aactttcgcc
      781 tttttaaaaa aatgtttcta atatctttaa aaattggaat attttggaaa acaatttgtt
      841 ttcaattggc aatttattaa ttctgagctt ctgagcgtaa attttttttt attttggagc
      901 aattctgaga tgtcttagtg catttttcgg gcatacataa aaaaaaattt atcttaataa
      961 taaaaacctt aaatttaggc ttcaccgtct ggaaattcaa ttttttcagc tgaaaaactt
     1021 tcaaaaaaaa aaaaattaaa ttttcacatt tattttctct ggaattttgg agcaatttca
     1081 aagctgtctt agtgcatttt tagtacacaa atattttttc atttttctta aattttaata
     1141 atctaagctt aaataaaggc ttcaccgtct gaaaatgcaa cttttttcag atgaaatatt
     1201 aaaaaaaaaa acctctttta ttaattttgt gctccaaact ttcccacggt acgcaaatca
     1261 tgtaatatgt acttttaaac attatttttc caatcaaaaa tatccaacag cttttcctta
     1321 caatttgaaa atttttatta aaatttcaaa aaaatttttt ttttttttta atttcgaatt
     1381 tttttaaact tgttgtaact tttttccctg tcggcgccaa ttaacaaaat acgtagaatt
     1441 ttgcgcttat tgttttgtta gttaacaaat ttttgacaac tttcttatca gctgagattt
     1501 aggtgtttca gttcgcaatc tgcgtctctc cttgctctcc tctattttaa aacccccata
     1561 actttgttct tgtgagagtt atcaaaaaat tgtcaactaa caaaatatag agcacaaaaa
     1621 tatgcacatt ttgtatgttg acaatttttt gatacctttt ttagttttcg agatatctta
     1681 gttttagttt agctgtacct ttttctggca aaaaacttcc aaatttcggc aatttttcgg
     1741 caaatttccc ggcaaatttc gccaagtaca tcgggctctc agacaagtgg tctgtgagtg
     1801 gaagtcgata aaagtagttt acgtgctctc cgtccccccg cttcttctcc gtctctgctt
     1861 ctatttctcc attgactagt cccactactt tttacggggc ggagggctct cttgcactcc
     1921 agcttcttct ctttttgtgc tcttttttgc ccgtatgtgg gtgctcattt atttttcttt
     1981 ggttttaaaa taaattcggt ttattaggct actcgtaaaa atgaagaaaa aacttcaaat
     2041 tcaaacgaca ttaaaaaaaa atttaataaa aattttttga attgctcaaa agccccacct
     2101 atatatatat atagctccgc ctacttctcg cacagacccc gcccactttt tgactccgcc
     2161 cccttattgg caaaagttct tgaaaaaaat aaaaataaaa attcaaattg aaacaatttt
     2221 tttttaatga agtagttttt ttctaaattg atgaaaattt ctaaacagtt tacaagcacc
     2281 gcccactttt tgactccgcc cctttgttgg aaaaattctt gtaaaaaaaa ataaaaaata
     2341 aaagcttctc tttcaaacga tttttagttg gtttttgtat gaaaagtttt tttcctaaaa
     2401 tagaaatgtc tgaataattt gcaagctccg cccttttctt acccagactc cgccccttta
     2461 ttgactaatt tttttttgtc catttacctg aagttcaaaa aaaaaattta cagtttcata
     2521 gagtaataat ttttcaaaaa tctctttttc caatgtttaa tgtgtcatta taaacccgcc
     2581 cagttttaag ctccgcccat ttaggccccg cccatttatg acccaaatat ttttttctct
     2641 attcacccgt ggatttccct ccaacaagtt accccaaaaa taaaaataaa aattcctgca
     2701 ggcgcccacc gggatccggc gcgcctctag aggatccccg ggattggcca aaggacccaa
     2761 aggtatgttt cgaatgatac taacataaca tagaacattt tcaggaggac ccttggctag
     2821 cgtcgacggt accaaaaatg gccgagggat ccgtcgcccg tcaaccagac cttcttacct
     2881 gcgagcacga ggagatccac cttgccggat ccatccaacc acacggagcc cttcttgtcg
     2941 tctccgagca cgaccaccgt gtcatccaag cctccgccaa cgccgccgag ttccttaacc
     3001 ttggatccgt ccttggagtc ccacttgccg agatcgacgg agaccttctt atcaaggtaa
     3061 gtttaaacat atatatacta actaaccctg attatttaaa ttttcagatc cttccacacc
     3121 ttgacccaac cgccgaggga atgccagtcg ccgtccgctg ccgcatcgga aacccatcca
     3181 ccgagtactg cggacttatg caccgcccac cagagggagg acttatcatc gagcttgagc
     3241 gtgccggacc atccatcgac ctttccggaa cccttgcccc agcccttgag cgcatccgta
     3301 ccgccggatc ccttcgtgcc ctttgcgacg acaccgtcct tcttttccag caatgcaccg
     3361 gatacgaccg tgtcatggtc taccgcttcg acgagcaagg acacggactt gtcttctccg
     3421 agtgccacgt cccaggactt gagtcctact tcggaaaccg ctacccatcc tccaccgtcc
     3481 cacaaatggc ccgtcaactt tacgtccgtc aacgtgtccg tgtccttgtc gacgtcacct
     3541 accaaccagt cccacttgag ccacgtcttt ccccacttac cggacgtgac cttgacatgt
     3601 ccggatgctt ccttcgttcc atgtccccat gccaccttca attccttaag gtaagtttaa
     3661 acagttcggt actaactaac catacatatt taaattttca ggacatggga gtccgtgcca
     3721 cccttgccgt ctcccttgtc gtcggaggaa agctttgggg acttgtcgtc tgccaccact
     3781 accttccacg cttcatccgc ttcgagcttc gtgccatctg caagcgtctt gccgagcgca
     3841 tcgccacccg tatcaccgcc cttgagtccg gatacgacat cggatccgga gagggacgcg
     3901 gatcccttct tacctgcgga gatgttgagg aaaacccagg accaatggag cgcccacaac
     3961 cagattctat gccacaagac ctttccgagg cccttaagga ggctaccaag gaggttcaca
     4021 cccaagctga gaacgctgag ttcatgcgca acttccaaaa gggacaagtt acccgtgatg
     4081 gattcaagct tgtcatggct tccctctacc acatctacgt cgctcttgag gaggagatcg
     4141 agcgcaacaa ggtaagttta aacatatata tactaactaa ccctgattat ttaaattttc
     4201 aggagtctcc agttttcgct ccagtttact tcccagaaga gctccaccgt aaggctgccc
     4261 ttgagcaaga ccttgccttc tggtatggac cacgctggca agaagttatc ccatacactc
     4321 cagctatgca acgctacgtt aagcgtcttc acgaggtcgg acgtaccgag ccagagcttc
     4381 ttgtcgctca cgcttacacc cgttaccttg gagacctttc cggaggacaa gtccttaagg
     4441 taagtttaaa cagttcggta ctaactaacc atacatattt aaattttcag aagatcgctc
     4501 aaaaggctct tgaccttcca tcttccggag agggacttgc cttcttcact ttcccaaaca
     4561 tcgcttctgc cactaagttc aagcaacttt accgctctcg tatgaactct cttgagatga
     4621 ccccagctgt ccgccaacgt gtcatcgagg aggctaaggt aagtttaaac atgattttac
     4681 taactaacta atctgattta aattttcaga ccgctttcct tcttaacatc caacttttcg
     4741 aagagcttca agagcttctt acccacgaca ccaaggacca atctccatcc cgtgccccag
     4801 gacttcgtca acgtgcctcc aacaaggtcc aagattccgc tccagttgag actccacgtg
     4861 gaaagccacc acttaacacc cgttcccaag ccccacttct tcgttgggtc cttaccctct
     4921 ccttccttgt cgccaccgtc gctgtcggac tttacgccat gtaagagctc cgcatcggcc
     4981 gctgtcatca gatcgccatc tcgcgcccgt gcctctgact tctaagtcca attactcttc
     5041 aacatcccta catgctcttt ctccctgtgc tcccaccccc tatttttgtt attatcaaaa
     5101 aaacttcttc ttaatttctt tgttttttag cttcttttaa gtcacctcta acaatgaaat
     5161 tgtgtagatt caaaaataga attaattcgt aataaaaagt cgaaaaaaat tgtgctccct
     5221 ccccccatta ataataattc tatcccaaaa tctacacaat gttctgtgta cacttcttat
     5281 gtttttttta cttctgataa attttttttg aaacatcata gaaaaaaccg cacacaaaat
     5341 accttatcat atgttacgtt tcagtttatg accgcaattt ttatttcttc gcacgtctgg
     5401 gcctctcatg acgtcaaatc atgctcatcg tgaaaaagtt ttggagtatt tttggaattt
     5461 ttcaatcaag tgaaagttta tgaaattaat tttcctgctt ttgctttttg ggggtttccc
     5521 ctattgtttg tcaagagttt cgaggacggc gtttttcttg ctaaaatcac aagtattgat
     5581 gagcacgatg caagaaagat cggaagaagg tttgggtttg aggctcagtg gaaggtgagt
     5641 agaagttgat aatttgaaag tggagtagtg tctatggggt ttttgcctta aatgacagaa
     5701 tacattccca atataccaaa cataactgtt tcctactagt cggccgtacg ggccctttcg
     5761 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5821 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5881 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     5941 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     6001 gccttaatta agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     6061 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     6121 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     6181 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     6241 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     6301 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     6361 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     6421 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     6481 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     6541 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     6601 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     6661 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     6721 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     6781 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6841 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6901 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6961 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     7021 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     7081 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     7141 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     7201 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     7261 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     7321 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     7381 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     7441 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     7501 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     7561 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7621 cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7681 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7741 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7801 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7861 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     7921 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     7981 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     8041 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     8101 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     8161 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     8221 t
//
