LOCUS       Exported                7321 bp ds-DNA     circular SYN 26-OCT-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7321)
  AUTHORS   Karl Clark
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Dec 8, 2021 from SnapGene 5.0.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7321
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1049..4259
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
                     /note="/locus_tag=source:recombinant plasmid"
                     /note="color: #ffc0cb"
     source          1193..1249
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
                     /note="/locus_tag=source:recombinant plasmid"
                     /note="color: #ffc0cb"
     rep_origin      complement(1..253)
                     /direction=LEFT
                     /locus_tag="ColE1 Ori"
                     /label=ColE1 Ori
                     /note="ColE1 Ori"
                     /note="color: #ffc0cb"
     primer_bind     241..266
                     /label=Ori_F1
                     /note="color: black; sequence: ggaagcggaacagatttaaatggtac; 
                     added: 2021-03-01"
     misc_feature    complement(267..457)
                     /locus_tag="ITR(R)"
                     /label=ITR(R)
                     /note="ITR(R)"
                     /note="Tol2 ITR(R)"
                     /note="color: #80ffff; direction: LEFT"
     misc_feature    487..506
                     /locus_tag="5' Universal Guide"
                     /label=5' Universal Guide
                     /note="color: #00ffff"
     misc_feature    506..509
                     /locus_tag="BfuAI site 1 overhang"
                     /label=BfuAI site 1 overhang
                     /note="color: #00ff00"
     misc_feature    514..519
                     /locus_tag="BfuA site 1"
                     /label=BfuA site 1
                     /note="color: #00ff00"
     promoter        520..641
                     /locus_tag="lacZ\promoter"
                     /label=lacZ promoter
                     /note="/vntifkey=30"
                     /note="color: #ffc0cb"
     misc_feature    642..917
                     /locus_tag="lacZ'"
                     /label=lacZ'
                     /note="/vntifkey=21"
                     /note="color: #ffc0cb"
     misc_feature    918..923
                     /locus_tag="BfuA site 2"
                     /label=BfuA site 2
                     /note="color: #00ffff"
     misc_feature    complement(928..938)
                     /locus_tag="TAL3"
                     /label=TAL3
                     /note="TAL3"
                     /note="color: #ffc0cb; direction: LEFT"
     misc_feature    928..931
                     /locus_tag="overhang5'3'"
                     /label=BfuAI site 2 overhang
                     /note="color: #00ffff"
     misc_feature    940..971
                     /locus_tag="rox(1)"
                     /label=rox
                     /note="color: #ff0080; direction: RIGHT"
     misc_feature    977..1010
                     /locus_tag="loxp"
                     /label=loxp
                     /note="color: #80ff80; direction: RIGHT"
     misc_feature    1015..1048
                     /locus_tag="lox2272"
                     /label=lox2272
                     /note="color: #8080ff; direction: RIGHT"
     intron          1060..1192
                     /locus_tag="BP/SA"
                     /label=BP/SA
                     /note="BP/SA"
                     /note="BP/SA from carp beta actin intron 1"
                     /note="color: #ffc0cb"
     primer_bind     complement(1168..1202)
                     /label=2A_SA_R0
                     /note="color: black; sequence: 
                     agttcgtggcctgtaaatgaatgagaaaaccggtt; added: 2021-03-01"
     primer_bind     1183..1210
                     /label=SA_2A_F0
                     /note="color: black; sequence: 
                     tcatttacaggccacgaacttctctctg; added: 2021-03-01"
     CDS             1193..1249
                     /codon_start=1
                     /locus_tag="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /product="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /label=2A peptide from porcine teschovirus-1
                     polyprotein
                     /label=2A peptide from porcine teschovirus-1 polyprotein
                     /note="P2A"
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /note="color: #00ff80"
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(1232..1258)
                     /label=RFP_2A_R
                     /note="color: black; sequence: ggaggaggcaggaccggggttttcttc;
                     added: 2021-01-08"
     primer_bind     1241..1267
                     /label=2A_RFP_F
                     /note="color: black; sequence: cccggtcctgcctcctccgaggacgtc;
                     added: 2021-01-08"
     CDS             1250..1924
                     /locus_tag="mRFP*"
                     /label=mRFP*
                     /note="mRFP*"
                     /note="color: #ffc0cb; direction: RIGHT"
     primer_bind     complement(1565..1602)
                     /label=mRFP_R
                     /note="color: black; sequence: 
                     atgaactcgccgtcctgcagggaggagtcctgggtcac; added: 2021-03-01"
     polyA_signal    1933..3108
                     /locus_tag="ocean pout terminator"
                     /label=ocean pout terminator
                     /note="ocean pout terminator"
                     /note="3'UTR, poly(A) signal, and putative boundary
                     element from ocean pout (FRM- Pat Gibbs)."
                     /note="color: #ffc0cb"
     primer_bind     3109..3149
                     /label=Primer 1
                     /note="color: black; sequence: 
                     gaagcttgagatcctagtcaaagctttcaaccactccaggc; added: 
                     2021-10-01"
     misc_feature    3135..3499
                     /locus_tag="He1a_promoter"
                     /label=He1a_promoter
                     /note="color: #ff0000"
     primer_bind     complement(3515..3534)
                     /label=RFP NLS R
                     /note="color: black; sequence: cttcttctttggagccatgg; added:
                     2021-10-04"
     misc_signal     3517..3546
                     /locus_tag="NLS"
                     /label=NLS
                     /note="NLS"
                     /note="color: #ffc0cb"
     CDS             3547..4248
                     /codon_start=1
                     /locus_tag="monomeric blue fluorescent protein"
                     /product="monomeric blue fluorescent protein"
                     /label=monomeric blue fluorescent protein
                     /note="TagBFP"
                     /note="mammalian codon-optimized"
                     /note="color: #ffc0cb"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    4260..4858
                     /locus_tag="Danio b-Actin 3' UTR"
                     /label=Danio b-Actin 3' UTR
                     /note="color: #0000ff"
     misc_feature    4839..5263
                     /locus_tag="Danio b-Actin poly(A) putative"
                     /label=Danio b-Actin poly(A) putative
                     /note="color: #004080"
     misc_feature    complement(5270..5303)
                     /locus_tag="loxp(1)"
                     /label=loxp
                     /note="color: #00ff80; direction: LEFT"
     misc_feature    complement(5309..5342)
                     /locus_tag="lox2272(1)"
                     /label=lox2272
                     /note="color: #800080; direction: LEFT"
     misc_feature    complement(5349..5380)
                     /locus_tag="rox"
                     /label=rox
                     /note="color: #ff00ff; direction: LEFT"
     misc_feature    5390..5392
                     /locus_tag="TAL3(1)"
                     /label=TAL3
                     /note="TAL3"
                     /note="color: #ffc0cb"
     misc_feature    5390..5392
                     /locus_tag="overhange3'5'"
                     /label=BspQI site 1 overhang
                     /note="color: #00ffff"
     misc_feature    5394..5400
                     /locus_tag="BspQ1 site1"
                     /label=BspQ1 site1
                     /note="color: #00ffff"
     intron          5409..5497
                     /locus_tag="Carp beta-actin intron 1 filler"
                     /label=Carp beta-actin intron 1 filler
                     /note="Carp beta-actin intron 1 filler"
                     /note="color: #ffc0cb"
     misc_feature    5500..5506
                     /locus_tag="BspQ1site2"
                     /label=BspQ1site2
                     /note="color: #00ffff"
     misc_feature    5508..5510
                     /locus_tag="BspQI site 2 overhang"
                     /label=BspQI site 2 overhang
                     /note="color: #00ffff"
     misc_feature    complement(5511..5530)
                     /locus_tag="Universal Guide 3'"
                     /label=Universal Guide 3'
                     /note="color: #00ff00; direction: LEFT"
     misc_feature    5572..5786
                     /locus_tag="ITR(L)"
                     /label=ITR(L)
                     /note="ITR(L)"
                     /note="Tol2 ITR(L)"
                     /note="color: #80ffff"
     CDS             5851..6660
                     /locus_tag="Kan(R)"
                     /label=Kan(R)
                     /note="Kan(R)"
                     /note="color: #ffc0cb; direction: RIGHT"
     rep_origin      complement(6693..7321)
                     /direction=LEFT
                     /locus_tag="ColE1 Ori(1)"
                     /label=ColE1 Ori
                     /note="ColE1 Ori"
                     /note="color: #ffc0cb"
ORIGIN
        1 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
       61 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
      121 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
      181 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
      241 ggaagcggaa cagatttaaa tggtaccgag cagaggtgta aaaagtactc aaaaatttta
      301 ctcaagtgaa agtacaagta cttagggaaa attttactca attaaaagta aaagtatctg
      361 gctagaatct tacttgagta aaagtaaaaa agtactccat taaaattgta cttgagtatt
      421 aaggaagtaa aagtaaaagc aagaaagaaa actagagtgg tctcccttta gtgagggtca
      481 attgatggga ggcgttcggg ccacagcgga cacgcaggtg cgcaacgcaa ttaatgtgag
      541 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
      601 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
      661 gcgcgcaatt aaccctcact aaagggaaca aaagctggag gacctcgagg gggggcccgg
      721 tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac
      781 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
      841 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
      901 gcctgaatgg cgaatggacc tgccacactt ccggccgcat aactttaaat aatgccaatt
      961 atttaaagtt actatcataa cttcgtatag catacattat acgaagttat caccataact
     1021 tcgtatagga tactttatac gaagttatcc taggcttacc acccactagc ggtcagactg
     1081 cagattgcag cacgaaacag gaagctgact ccacatggtc acatgctcac tgaagtgttg
     1141 acttccctga cagctgtgca ctttctaaac cggttttctc attcatttac aggccacgaa
     1201 cttctctctg ttaaagcaag caggagacgt ggaagaaaac cccggtcctg cctcctccga
     1261 ggacgtcatc aaggagttca tgcgcttcaa ggtgcgcatg gagggctccg tgaacggcca
     1321 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     1381 gctgaaggtg accaagggcg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
     1441 ccagtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
     1501 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
     1561 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
     1621 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc
     1681 ctccaccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agatgaggct
     1741 gaagctgaag gacggcggcc actacgacgc cgaggtcaag accacctaca tggccaagaa
     1801 gcccgtgcag ctgcccggcg cctacaagac cgacatcaag ctggacatca cctcccacaa
     1861 cgaggactac accatcgtgg aacagtacga gcgcgccgag ggccgccact ccaccggagc
     1921 ctaagactag taattaagtc tcagccaccg ttaactgaac atgtcaaaac ctgtggagac
     1981 tgttgagatt tgatgttctg aaaagataaa gcctataaat aaaatgttgc ccaaatttcc
     2041 tgcctgatgt ttttctttgt ctttgctaca tggctttgct gctcggatcg gctcactctg
     2101 tgtatgccac gttcactttg tactctcctt ctcacggtag gtttattatt tttagatgtg
     2161 cagttagttt ctgtgaaata acacaccaca cactgatatt gtctgtgcat tgacttggtg
     2221 agtgcacatt gtttttgatc ttgacatatt tatatttgat tgatcaggtg aactgtgtga
     2281 atctaaagtg ctccatacag atgttctgca ttgaaaatat tctcatttta ttagtggaag
     2341 tgagtgtatg ccacatccaa tcaatttcag caaacacccc agtatgattt aatgcaaaaa
     2401 aatgaaggta tcaaacacgc attactactt tgcagttaaa tatttaacat ttattccaac
     2461 acgaaaaaaa gcagtaaata acactttgac aaacacgtca ggacatctta tttttgtcac
     2521 cctcacaggc aatttagtat aatatattat atatatatat atatcatata ataatattca
     2581 gtataatata tatatatata tcatattata atattcagta taatataaaa cacaaacaca
     2641 tatatgtata atataatata acatttttat ttattgagat gcctctatgg accgtgttat
     2701 aagaagtaaa gatcaggaga agtaaacatg aagtgtaatt atgaatactg atgttaaatt
     2761 aagctatgat gagttttcac tgttaattta ccatctcaat taaatgttga tgcctccatg
     2821 accaagttaa gcagatgaga ctgagacaac tgtagaagac aagatgttca ctttgctgaa
     2881 tatagctggc ttgacagtta tctatgactc tataaatata tatatatttt tttttttata
     2941 aaatgattta tttataacta tatatccatt tctcagacag gtgcttcata tccctcactc
     3001 ccgtagctgt ccatgctgga tctgtccccg ttgtttttaa aaagctaaat aagttattaa
     3061 catgactgca tccagcgagc caaacctgtc tggtgtacag ctaccagaga agcttgagat
     3121 cctagtcaaa gctttcaacc actccaggca tagctagagc cactgatgca tctatagagc
     3181 tgaagtaaag atagggttga tgcaaatcga tttagaattt tcaatgtaga aaaaacaaaa
     3241 taaaaagtct attcacttga tggatagatt agtcaaacaa acatagagta ggcacttccc
     3301 ttgactctgt aaacttttgg atttaacaga ttgtgcactc cttttagtta aacacattag
     3361 tccctgaaaa ctaagaccct aaccataatc ccattgtaga cccacctact cgggtgaata
     3421 taaagctcgc acagatcatc tcacagacat cagaccttgg caaaggagca tcctgaacat
     3481 ctctacacac tgaggcaacg aattcacctc gagaccatgg ctccaaagaa gaagcgtaag
     3541 gtaaccatga gcgagctgat taaggagaac atgcacatga agctgtacat ggagggcacc
     3601 gtggacaacc atcacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc
     3661 cagaccatga gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg
     3721 gctactagct tcctctacgg cagcaagacc ttcatcaacc acacccaggg catccccgac
     3781 ttcttcaagc agtccttccc tgagggcttc acatgggaga gagtcaccac atacgaagac
     3841 gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac
     3901 gtcaagatca gaggggtgaa cttcacatcc aacggccctg tgatgcagaa gaaaacactc
     3961 ggctgggagg ccttcaccga gacgctgtac cccgctgacg gcggcctgga aggcagaaac
     4021 gacatggccc tgaagctcgt gggcgggagc catctgatcg caaacatcaa gaccacatat
     4081 agatccaaga aacccgctaa gaacctcaag atgcctggcg tctactatgt ggactacaga
     4141 ctggaaagaa tcaaggaggc caacaacgag acctacgtcg agcagcacga ggtggcagtg
     4201 gccagatact gcgacctccc tagcaaactg gggcacaagc tcaattaaag ctgagatcta
     4261 cggactgtta ccacttcacg ccgactcaaa ctgcgcagag aaacttcaaa cgacaacatt
     4321 ggcatggctt ttgttatttt tggcgcttga ctcaggatct aaaaactgga acggtgaagg
     4381 tgacggcaat gttttttggc aaataagcat ccccgaagtt ctacaatgca tctgaggact
     4441 ccatgttttg ttttctttag tcattccaaa tgtttgttaa atgcattgtt ccgaaactta
     4501 tttgcctcta tgaaggctgc ccagtaattg ggagcatact taacattgta gtattgtatg
     4561 taaattatgt aacaaaacaa tgtctgggtt tttgtacttt tcagccttaa aatcttgggt
     4621 ttattttttt ttctttgttc caaaaaaacg aagctttacc attcaagatg taaaggtttc
     4681 cattccccct gggcatattg taaaagctgt gtggaacgtg gcggtgccag acatttggtg
     4741 gggccaacct gtacactgac taattcaatt ccaataaaag tgcacatgta agacatccta
     4801 cccctgtctc atttcctgtc ttggctctga gtggcttcaa taaaaatggc ttttgtgaag
     4861 ttgcaccatg ttaattacat ttcagcactt tcacaccagg tttttgacta atttgaattt
     4921 atcgaagttt gacctttttt tttttttttc ttcaagctaa tttgaggtta tcaaaccatt
     4981 tcattggagg aaacttaatg tttaaaagca taaattcgaa acatgattca tttatttaac
     5041 tgcttaatgg tgaaacaaac ttcaagtaat acaccaagtg ttccctcata tgaaggtagc
     5101 taactgcaaa cggccttaac tttccatctt aagttgagct acataaaggc tttaacagga
     5161 aggaactgcc agtttaaaag tcaaaccacc atgactgata taaattcttg tccatataac
     5221 cttgcgtttc acacaatata gctatctact gctaaataaa ttagagctca taacttcgta
     5281 taatgtatgc tatacgaagt tatgttagat aacttcgtat aaagtatcct atacgaagtt
     5341 atgcgagcta actttaaata attggcatta tttaaagtta tgaatcgata agtgaagagc
     5401 tatccggata tatatgtatg ttatcataac tgtactggct atgtcaggtg gtaatgactg
     5461 taacgttacg ttactcgttg taggcacgac attgaatatg ctcttctccg ctgtggcccg
     5521 aacgcctccc gctagcgatg ggatctaata cgactcacta tagggagacc aagggaaaat
     5581 agaatgaagt gatctccaaa aaataagtac tttttgactg taaataaaat tgtaaggagt
     5641 aaaaagtact tttttttcta aaaaaatgta attaagtaaa agtaaaagta ttgattttta
     5701 attgtactca agtaaagtaa aaatccccaa aaataatact taagtacagt aatcaagtaa
     5761 aattactcaa gtactttaca cctctgggat ccagctgcat catgaacaat aaaactgtct
     5821 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg
     5881 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     5941 gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg
     6001 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     6061 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     6121 gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa
     6181 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     6241 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg
     6301 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     6361 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca
     6421 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     6481 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     6541 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     6601 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     6661 tcagaattgg ttaattggtt gtaacactgg caaaaaggat ctaggtgaag atcctttttg
     6721 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6781 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6841 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6901 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6961 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     7021 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     7081 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     7141 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     7201 aaagcgccac ggttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     7261 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     7321 t
//
