LOCUS       Exported                8239 bp ds-DNA     circular SYN 27-MAY-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8239)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jan 20, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8239
                     /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=0507-624-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..3887
                     /codon_start=1
                     /label=wmRhubarb713
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKVSLNI
                     YILTNPDYLNFQDAGFIGSWHRHDQLIFLELEPPQRDVAEPQAFFRHTNSAIRRLQAAE
                     TLESACAAAAQEVRKVSLNSSVLTNHTYLNFQITGFDRVMIYRFASDFSGEVIAEDRCA
                     EVESNLGLHYPASTVPAQARRLYTINPVRIIPDINYRPVPVTPDLNPVTGRPIDLSFAI
                     LRSVSPVHLEFMRNIGMHGTMSISILRGERLWGLIVCHHRTPYYVDLDGRQACELVAQV
                     LARAIGVMEE"
     primer_bind     complement(2999..3009)
                     /label=0507-624-R
     misc_feature    3135..3185
                     /label=intron-1
     misc_feature    3372..3422
                     /label=intron-2
     primer_bind     complement(3866..3908)
                     /label=0507-624-R
     CDS             3888..3908
                     /codon_start=1
                     /label=Linker
                     /translation="GYDIGSG"
     CDS             3909..3962
                     /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             3963..4982
                     /codon_start=1
                     /label=HO1
                     /translation="MERPQPDSMPQDLSEALKEATKEVHTQAENAEFMRNFQKGQVTRD
                     GFKLVMASLYHIYVALEEEIERNKVSLNIYILTNPDYLNFQESPVFAPVYFPEELHRKA
                     ALEQDLAFWYGPRWQEVIPYTPAMQRYVKRLHEVGRTEPELLVAHAYTRYLGDLSGGQV
                     LKVSLNSSVLTNHTYLNFQKIAQKALDLPSSGEGLAFFTFPNIASATKFKQLYRSRMNS
                     LEMTPAVRQRVIEEAKVSLNMILLTN*SDLNFQTAFLLNIQLFEELQELLTHDTKDQSP
                     SRAPGLRQRASNKVQDSAPVETPRGKPPLNTRSQAPLLRWVLTLSFLVATVAVGLYAM"
     misc_feature    4170..4220
                     /label=Intron-1
     misc_feature    4458..4508
                     /label=Intron-2
     misc_feature    4677..4727
                     /label=Intron-3
     primer_bind     complement(5024..5040)
                     /label=pSF11-Rv-Seq
     promoter        6118..6222
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6223..7083
                     /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             6421..7080
                     /label=AmpR
     rep_origin      7178..7860
                     /label=ColE1 origin
     rep_origin      7254..7842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8130..8151
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8166..8196
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     misc_binding    8200..8222
                     /label=LacO
     protein_bind    8204..8220
                     /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(8228..8239,1..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 gcgacgacga gccaatccac atcccaggag ccatccagcc acacggactt cttcttgccc
     2941 ttgccgccga catgaccatc gtcgccggat ccgacaacct tccagagctt accggacttg
     3001 ccatcggagc ccttatcgga cgttccgccg ccgacgtctt cgactccgag acccacaacc
     3061 gccttaccat cgcccttgcc gagccaggag ccgccgtcgg agccccaatc accgtcggat
     3121 tcaccatgcg caaggtaagt ttaaacatat atatactaac taaccctgat tatttaaatt
     3181 ttcaggacgc cggattcatc ggatcctggc accgtcacga ccaacttatc ttccttgagc
     3241 ttgagccacc acaacgcgac gtcgccgagc cacaagcctt cttccgccac accaactccg
     3301 ccatccgtcg tcttcaagcc gccgagaccc ttgagtccgc ctgcgccgcc gccgcccaag
     3361 aggtccgcaa ggtaagttta aacagttcgg tactaactaa ccatacatat ttaaattttc
     3421 agatcaccgg attcgaccgc gtcatgatct accgtttcgc ctccgacttc tccggagagg
     3481 tcatcgccga ggaccgttgc gccgaggtcg agtccaacct tggacttcac tacccagcct
     3541 ccaccgtccc agcccaagcc cgccgccttt acaccatcaa cccagtccgc atcatcccag
     3601 acatcaacta ccgcccagtc ccagtcaccc cagaccttaa cccagtcacc ggacgcccaa
     3661 tcgacctttc cttcgccatc cttcgttccg tctccccagt ccaccttgag ttcatgcgca
     3721 acatcggaat gcacggaacc atgtccatct ccatccttcg cggagagcgt ctttggggac
     3781 ttatcgtctg ccaccaccgc accccatact acgtcgacct tgacggacgt caagcctgcg
     3841 agcttgtcgc ccaagtcctt gcccgtgcca tcggagtcat ggaggaggga tacgacatcg
     3901 gatccggaga gggacgcgga tcccttctta cctgcggaga tgttgaggaa aacccaggac
     3961 caatggagcg cccacaacca gattctatgc cacaagacct ttccgaggcc cttaaggagg
     4021 ctaccaagga ggttcacacc caagctgaga acgctgagtt catgcgcaac ttccaaaagg
     4081 gacaagttac ccgtgatgga ttcaagcttg tcatggcttc cctctaccac atctacgtcg
     4141 ctcttgagga ggagatcgag cgcaacaagg taagtttaaa catatatata ctaactaacc
     4201 ctgattattt aaattttcag gagtctccag ttttcgctcc agtttacttc ccagaagagc
     4261 tccaccgtaa ggctgccctt gagcaagacc ttgccttctg gtatggacca cgctggcaag
     4321 aagttatccc atacactcca gctatgcaac gctacgttaa gcgtcttcac gaggtcggac
     4381 gtaccgagcc agagcttctt gtcgctcacg cttacacccg ttaccttgga gacctttccg
     4441 gaggacaagt ccttaaggta agtttaaaca gttcggtact aactaaccat acatatttaa
     4501 attttcagaa gatcgctcaa aaggctcttg accttccatc ttccggagag ggacttgcct
     4561 tcttcacttt cccaaacatc gcttctgcca ctaagttcaa gcaactttac cgctctcgta
     4621 tgaactctct tgagatgacc ccagctgtcc gccaacgtgt catcgaggag gctaaggtaa
     4681 gtttaaacat gattttacta actaactaat ctgatttaaa ttttcagacc gctttccttc
     4741 ttaacatcca acttttcgaa gagcttcaag agcttcttac ccacgacacc aaggaccaat
     4801 ctccatcccg tgccccagga cttcgtcaac gtgcctccaa caaggtccaa gattccgctc
     4861 cagttgagac tccacgtgga aagccaccac ttaacacccg ttcccaagcc ccacttcttc
     4921 gttgggtcct taccctctcc ttccttgtcg ccaccgtcgc tgtcggactt tacgccatgt
     4981 aagagctccg catcggccgc tgtcatcaga tcgccatctc gcgcccgtgc ctctgacttc
     5041 taagtccaat tactcttcaa catccctaca tgctctttct ccctgtgctc ccacccccta
     5101 tttttgttat tatcaaaaaa acttcttctt aatttctttg ttttttagct tcttttaagt
     5161 cacctctaac aatgaaattg tgtagattca aaaatagaat taattcgtaa taaaaagtcg
     5221 aaaaaaattg tgctccctcc ccccattaat aataattcta tcccaaaatc tacacaatgt
     5281 tctgtgtaca cttcttatgt tttttttact tctgataaat tttttttgaa acatcataga
     5341 aaaaaccgca cacaaaatac cttatcatat gttacgtttc agtttatgac cgcaattttt
     5401 atttcttcgc acgtctgggc ctctcatgac gtcaaatcat gctcatcgtg aaaaagtttt
     5461 ggagtatttt tggaattttt caatcaagtg aaagtttatg aaattaattt tcctgctttt
     5521 gctttttggg ggtttcccct attgtttgtc aagagtttcg aggacggcgt ttttcttgct
     5581 aaaatcacaa gtattgatga gcacgatgca agaaagatcg gaagaaggtt tgggtttgag
     5641 gctcagtgga aggtgagtag aagttgataa tttgaaagtg gagtagtgtc tatggggttt
     5701 ttgccttaaa tgacagaata cattcccaat ataccaaaca taactgtttc ctactagtcg
     5761 gccgtacggg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     5821 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     5881 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     5941 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     6001 aaaataccgc atcaggcggc cttaattaag ggcctcgtga tacgcctatt tttataggtt
     6061 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     6121 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     6181 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     6241 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     6301 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     6361 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     6421 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     6481 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     6541 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     6601 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     6661 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     6721 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     6781 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     6841 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     6901 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     6961 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     7021 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     7081 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     7141 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     7201 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     7261 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     7321 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     7381 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     7441 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     7501 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     7561 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     7621 gaactgagat acctacagcg tgagcattga gaaagcgcca cgcttcccga agggagaaag
     7681 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     7741 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     7801 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     7861 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     7921 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     7981 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     8041 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     8101 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     8161 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     8221 tttcacacag gaaacagct
//
