LOCUS       Exported                8224 bp ds-DNA     circular SYN 13-SEP-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8224)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jan 20, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8224
                     /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..2861
                     /label=0914-693-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..3872
                     /codon_start=1
                     /label=wmiRFP670-2
                     /translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
                     AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
                     RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKVSLNIYILTNPDYLNFQELKSLEEMA
                     ARVPRYLQAMLGYHRVMLYRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLY
                     LKVSLNSSVLTNHTYLNFQNAIRVVSDSRGISSRIVPEHDASGAALDLSFAHLRSISPC
                     HLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYEPRAVPMAQRVAAKRFAERLSTHFTA
                     AHHQR"
     misc_feature    3249..3299
                     /label=intron-1
     misc_feature    3510..3560
                     /label=intron-2
     CDS             3873..3893
                     /codon_start=1
                     /label=Linker
                     /translation="GYDIGSG"
     CDS             3894..3947
                     /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             3948..4967
                     /codon_start=1
                     /label=HO1
                     /translation="MERPQPDSMPQDLSEALKEATKEVHTQAENAEFMRNFQKGQVTRD
                     GFKLVMASLYHIYVALEEEIERNKVSLNIYILTNPDYLNFQESPVFAPVYFPEELHRKA
                     ALEQDLAFWYGPRWQEVIPYTPAMQRYVKRLHEVGRTEPELLVAHAYTRYLGDLSGGQV
                     LKVSLNSSVLTNHTYLNFQKIAQKALDLPSSGEGLAFFTFPNIASATKFKQLYRSRMNS
                     LEMTPAVRQRVIEEAKVSLNMILLTN*SDLNFQTAFLLNIQLFEELQELLTHDTKDQSP
                     SRAPGLRQRASNKVQDSAPVETPRGKPPLNTRSQAPLLRWVLTLSFLVATVAVGLYAM"
     misc_feature    4155..4205
                     /label=Intron-1
     misc_feature    4443..4493
                     /label=Intron-2
     misc_feature    4662..4712
                     /label=Intron-3
     primer_bind     complement(4944..4993)
                     /label=0914-693-R
     primer_bind     complement(5009..5025)
                     /label=pSF11-Rv-Seq
     promoter        6103..6207
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6208..7068
                     /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             6406..7065
                     /label=AmpR
     rep_origin      7163..7845
                     /label=ColE1 origin
     rep_origin      7239..7827
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8115..8136
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8151..8181
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     misc_binding    8185..8207
                     /label=LacO
     protein_bind    8189..8205
                     /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(8213..8224,1..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 gcccgtaagg tcgaccttac ctcctgcgac cgcgagccaa
     2881 tccacatccc aggatccatc cagccatgcg gatgccttct tgcctgcgac gcccaagccg
     2941 tccgcatcac ccgcatcacc gagaacgccg gagccttctt cggacgcgag accccacgtg
     3001 tcggagagct tcttgccgac tacttcggag agaccgaggc ccacgccctt cgtaacgccc
     3061 ttgcccaatc ctccgaccca aagcgtccag cccttatctt cggatggcgc gacggactta
     3121 ccggacgcac cttcgacatc tcccttcacc gtcacgacgg aacctccatc atcgagttcg
     3181 agccagccgc cgccgagcaa gccgacaacc cacttcgcct tacccgccaa atcatcgccc
     3241 gcaccaaggt aagtttaaac atatatatac taactaaccc tgattattta aattttcagg
     3301 agcttaagtc ccttgaggag atggccgccc gtgtcccacg ttaccttcaa gccatgcttg
     3361 gataccaccg tgtcatgctt taccgtttcg ccgacgacgg atccggaatg gtcatcggag
     3421 aggccaagcg ttccgacctt gagtccttcc ttggacaaca cttcccagcc tcccttgtcc
     3481 cacaacaagc ccgtcttctt taccttaagg taagtttaaa cagttcggta ctaactaacc
     3541 atacatattt aaattttcag aacgccatcc gtgtcgtctc cgactcccgt ggaatctcct
     3601 cccgtatcgt cccagagcac gacgcctccg gagccgccct tgacctttcc ttcgcccacc
     3661 ttcgttccat ctccccatgc caccttgagt tccttcgcaa catgggagtc tccgcctcca
     3721 tgtccctttc catcatcatc gacggaaccc tttggggact tatcatctgc caccactacg
     3781 agccacgtgc cgtcccaatg gcccaacgtg tcgccgccaa gcgtttcgcc gagcgtcttt
     3841 ccacccactt caccgccgcc caccaccaac gtggatacga catcggatcc ggagagggac
     3901 gcggatccct tcttacctgc ggagatgttg aggaaaaccc aggaccaatg gagcgcccac
     3961 aaccagattc tatgccacaa gacctttccg aggcccttaa ggaggctacc aaggaggttc
     4021 acacccaagc tgagaacgct gagttcatgc gcaacttcca aaagggacaa gttacccgtg
     4081 atggattcaa gcttgtcatg gcttccctct accacatcta cgtcgctctt gaggaggaga
     4141 tcgagcgcaa caaggtaagt ttaaacatat atatactaac taaccctgat tatttaaatt
     4201 ttcaggagtc tccagttttc gctccagttt acttcccaga agagctccac cgtaaggctg
     4261 cccttgagca agaccttgcc ttctggtatg gaccacgctg gcaagaagtt atcccataca
     4321 ctccagctat gcaacgctac gttaagcgtc ttcacgaggt cggacgtacc gagccagagc
     4381 ttcttgtcgc tcacgcttac acccgttacc ttggagacct ttccggagga caagtcctta
     4441 aggtaagttt aaacagttcg gtactaacta accatacata tttaaatttt cagaagatcg
     4501 ctcaaaaggc tcttgacctt ccatcttccg gagagggact tgccttcttc actttcccaa
     4561 acatcgcttc tgccactaag ttcaagcaac tttaccgctc tcgtatgaac tctcttgaga
     4621 tgaccccagc tgtccgccaa cgtgtcatcg aggaggctaa ggtaagttta aacatgattt
     4681 tactaactaa ctaatctgat ttaaattttc agaccgcttt ccttcttaac atccaacttt
     4741 tcgaagagct tcaagagctt cttacccacg acaccaagga ccaatctcca tcccgtgccc
     4801 caggacttcg tcaacgtgcc tccaacaagg tccaagattc cgctccagtt gagactccac
     4861 gtggaaagcc accacttaac acccgttccc aagccccact tcttcgttgg gtccttaccc
     4921 tctccttcct tgtcgccacc gtcgctgtcg gactttacgc catgtaagag ctccgcatcg
     4981 gccgctgtca tcagatcgcc atctcgcgcc cgtgcctctg acttctaagt ccaattactc
     5041 ttcaacatcc ctacatgctc tttctccctg tgctcccacc ccctattttt gttattatca
     5101 aaaaaacttc ttcttaattt ctttgttttt tagcttcttt taagtcacct ctaacaatga
     5161 aattgtgtag attcaaaaat agaattaatt cgtaataaaa agtcgaaaaa aattgtgctc
     5221 cctcccccca ttaataataa ttctatccca aaatctacac aatgttctgt gtacacttct
     5281 tatgtttttt ttacttctga taaatttttt ttgaaacatc atagaaaaaa ccgcacacaa
     5341 aataccttat catatgttac gtttcagttt atgaccgcaa tttttatttc ttcgcacgtc
     5401 tgggcctctc atgacgtcaa atcatgctca tcgtgaaaaa gttttggagt atttttggaa
     5461 tttttcaatc aagtgaaagt ttatgaaatt aattttcctg cttttgcttt ttgggggttt
     5521 cccctattgt ttgtcaagag tttcgaggac ggcgtttttc ttgctaaaat cacaagtatt
     5581 gatgagcacg atgcaagaaa gatcggaaga aggtttgggt ttgaggctca gtggaaggtg
     5641 agtagaagtt gataatttga aagtggagta gtgtctatgg ggtttttgcc ttaaatgaca
     5701 gaatacattc ccaatatacc aaacataact gtttcctact agtcggccgt acgggccctt
     5761 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     5821 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     5881 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     5941 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     6001 gcggccttaa ttaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     6061 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     6121 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     6181 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     6241 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     6301 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     6361 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     6421 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6481 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6541 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6601 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6661 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6721 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6781 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6841 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6901 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6961 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     7021 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     7081 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     7141 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     7201 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     7261 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     7321 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     7381 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7441 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7501 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7561 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7621 cagcgtgagc attgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7681 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7741 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7801 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7861 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     7921 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     7981 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     8041 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     8101 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     8161 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     8221 agct
//
