LOCUS       Exported                7587 bp ds-DNA     circular SYN 25-APR-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7587)
  AUTHORS   Karl Clark
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jul 8, 2021 from SnapGene 5.0.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7587
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1049..4525
                     /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    complement(506..509)
                     /locus_tag="BfuAI site 1 overhang"
                     /label=BfuAI site 1 overhang
                     /note="color: #00ff00; direction: LEFT"
     misc_feature    complement(514..519)
                     /locus_tag="BfuA site 1"
                     /label=BfuA site 1
                     /note="color: #00ff00; direction: LEFT"
     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 5'3'
                     /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"
     misc_feature    3152..3512
                     /locus_tag="gamma-crystallin"
                     /label=gamma-crystallin
                     /note="gamma-crystallin"
                     /note="Xenopus"
                     /note="color: #ffc0cb"
     intron          3565..3762
                     /locus_tag="mIntron"
                     /label=mIntron
                     /note="mIntron"
                     /note="carp bactin shortened intron I"
                     /note="color: #ffc0cb"
     primer_bind     complement(3733..3757)
                     /label=2A_SA_R0
                     /note="color: black; sequence: 
                     agttcgtggcctgtaaatgaatgagaaaaccggtt; added: 2021-03-01"
     misc_signal     3783..3812
                     /locus_tag="NLS"
                     /label=NLS
                     /note="NLS"
                     /note="color: #ffc0cb"
     CDS             3813..4514
                     /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    4526..5124
                     /locus_tag="Danio b-Actin 3' UTR"
                     /label=Danio b-Actin 3' UTR
                     /note="color: #0000ff"
     misc_feature    5105..5529
                     /locus_tag="Danio b-Actin poly(A) putative"
                     /label=Danio b-Actin poly(A) putative
                     /note="color: #004080"
     misc_feature    complement(5536..5569)
                     /locus_tag="loxp(1)"
                     /label=loxp
                     /note="color: #00ff80; direction: LEFT"
     misc_feature    complement(5575..5608)
                     /locus_tag="lox2272(1)"
                     /label=lox2272
                     /note="color: #800080; direction: LEFT"
     misc_feature    complement(5615..5646)
                     /locus_tag="rox"
                     /label=rox
                     /note="color: #ff00ff; direction: LEFT"
     misc_feature    5656..5658
                     /locus_tag="TAL3(1)"
                     /label=TAL3
                     /note="TAL3"
                     /note="color: #ffc0cb"
     misc_feature    5656..5658
                     /locus_tag="overhange3'5'"
                     /label=BspQI site 1 overhang
                     /note="color: #00ffff"
     misc_feature    5660..5666
                     /locus_tag="BspQ1 site1"
                     /label=BspQ1 site1
                     /note="color: #00ffff"
     intron          5675..5763
                     /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    5766..5772
                     /locus_tag="BspQ1site2"
                     /label=BspQ1site2
                     /note="color: #00ffff"
     misc_feature    5774..5776
                     /locus_tag="BspQI site 2 overhang"
                     /label=BspQI site 2 overhang
                     /note="color: #00ffff"
     misc_feature    complement(5777..5796)
                     /locus_tag="Universal Guide 3'"
                     /label=Universal Guide 3'
                     /note="color: #00ff00; direction: LEFT"
     misc_feature    5838..6052
                     /locus_tag="ITR(L)"
                     /label=ITR(L)
                     /note="ITR(L)"
                     /note="Tol2 ITR(L)"
                     /note="color: #80ffff"
     CDS             6117..6926
                     /locus_tag="Kan(R)"
                     /label=Kan(R)
                     /note="Kan(R)"
                     /note="color: #ffc0cb; direction: RIGHT"
     rep_origin      complement(6959..7587)
                     /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 cctagtcacc ggcgttccta gttctttaaa ctgttacaaa gtgtttggac cagcaactga
     3181 attgcactgt aaattaccac atcagggtct ccggggtcaa cgctgctaag caatggactg
     3241 gaaaatttgt gacattgagt ttttggaata actgggaatg ctggataact ctttgggtaa
     3301 agctatggaa caaaagtacc agcatgaact aagaaatgac tgtcacagca caacatcaaa
     3361 cacaatgcaa gacaaaggtt ctctgctttt gccttacttg acagaaaaag taagctctag
     3421 gttcatcgca tgcagacagc agcaatacat gacatatata aaggctggct cttttctccc
     3481 ttccattgaa ctgaaacttc cactcagtca gacttgcggg ggatccacag cctgactaac
     3541 tcgacaaacc cccccaaacc taaggtgagt tgatctttaa gctttttaca ttttcagctc
     3601 gcatatatca attcgcgcgc attgtcacac tagcacccac tagcggtcag actgcagaca
     3661 aacaggaagc tgactccaca tggtcacatg ctcactgaag tgttgacttc cctgacagct
     3721 gtgcactttc taaaccggtt ttctcattca tttacagttc agccggaatt cacctcgaga
     3781 ccatggctcc aaagaagaag cgtaaggtaa ccatgagcga gctgattaag gagaacatgc
     3841 acatgaagct gtacatggag ggcaccgtgg acaaccatca cttcaagtgc acatccgagg
     3901 gcgaaggcaa gccctacgag ggcacccaga ccatgagaat caaggtggtc gagggcggcc
     3961 ctctcccctt cgccttcgac atcctggcta ctagcttcct ctacggcagc aagaccttca
     4021 tcaaccacac ccagggcatc cccgacttct tcaagcagtc cttccctgag ggcttcacat
     4081 gggagagagt caccacatac gaagacgggg gcgtgctgac cgctacccag gacaccagcc
     4141 tccaggacgg ctgcctcatc tacaacgtca agatcagagg ggtgaacttc acatccaacg
     4201 gccctgtgat gcagaagaaa acactcggct gggaggcctt caccgagacg ctgtaccccg
     4261 ctgacggcgg cctggaaggc agaaacgaca tggccctgaa gctcgtgggc gggagccatc
     4321 tgatcgcaaa catcaagacc acatatagat ccaagaaacc cgctaagaac ctcaagatgc
     4381 ctggcgtcta ctatgtggac tacagactgg aaagaatcaa ggaggccaac aacgagacct
     4441 acgtcgagca gcacgaggtg gcagtggcca gatactgcga cctccctagc aaactggggc
     4501 acaagctcaa ttaaagctga gatctacgga ctgttaccac ttcacgccga ctcaaactgc
     4561 gcagagaaac ttcaaacgac aacattggca tggcttttgt tatttttggc gcttgactca
     4621 ggatctaaaa actggaacgg tgaaggtgac ggcaatgttt tttggcaaat aagcatcccc
     4681 gaagttctac aatgcatctg aggactccat gttttgtttt ctttagtcat tccaaatgtt
     4741 tgttaaatgc attgttccga aacttatttg cctctatgaa ggctgcccag taattgggag
     4801 catacttaac attgtagtat tgtatgtaaa ttatgtaaca aaacaatgtc tgggtttttg
     4861 tacttttcag ccttaaaatc ttgggtttat ttttttttct ttgttccaaa aaaacgaagc
     4921 tttaccattc aagatgtaaa ggtttccatt ccccctgggc atattgtaaa agctgtgtgg
     4981 aacgtggcgg tgccagacat ttggtggggc caacctgtac actgactaat tcaattccaa
     5041 taaaagtgca catgtaagac atcctacccc tgtctcattt cctgtcttgg ctctgagtgg
     5101 cttcaataaa aatggctttt gtgaagttgc accatgttaa ttacatttca gcactttcac
     5161 accaggtttt tgactaattt gaatttatcg aagtttgacc tttttttttt tttttcttca
     5221 agctaatttg aggttatcaa accatttcat tggaggaaac ttaatgttta aaagcataaa
     5281 ttcgaaacat gattcattta tttaactgct taatggtgaa acaaacttca agtaatacac
     5341 caagtgttcc ctcatatgaa ggtagctaac tgcaaacggc cttaactttc catcttaagt
     5401 tgagctacat aaaggcttta acaggaagga actgccagtt taaaagtcaa accaccatga
     5461 ctgatataaa ttcttgtcca tataaccttg cgtttcacac aatatagcta tctactgcta
     5521 aataaattag agctcataac ttcgtataat gtatgctata cgaagttatg ttagataact
     5581 tcgtataaag tatcctatac gaagttatgc gagctaactt taaataattg gcattattta
     5641 aagttatgaa tcgataagtg aagagctatc cggatatata tgtatgttat cataactgta
     5701 ctggctatgt caggtggtaa tgactgtaac gttacgttac tcgttgtagg cacgacattg
     5761 aatatgctct tctccgctgt ggcccgaacg cctcccgcta gcgatgggat ctaatacgac
     5821 tcactatagg gagaccaagg gaaaatagaa tgaagtgatc tccaaaaaat aagtactttt
     5881 tgactgtaaa taaaattgta aggagtaaaa agtacttttt tttctaaaaa aatgtaatta
     5941 agtaaaagta aaagtattga tttttaattg tactcaagta aagtaaaaat ccccaaaaat
     6001 aatacttaag tacagtaatc aagtaaaatt actcaagtac tttacacctc tgggatccag
     6061 ctgcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
     6121 gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat
     6181 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt
     6241 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
     6301 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
     6361 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggaa
     6421 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     6481 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     6541 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     6601 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     6661 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     6721 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     6781 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     6841 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     6901 atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaaa
     6961 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     7021 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     7081 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     7141 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     7201 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     7261 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     7321 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     7381 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     7441 gagataccta cagcgtgagc tatgagaaag cgccacggtt cccgaaggga gaaaggcgga
     7501 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     7561 aaacgcctgg tatctttata gtcctgt
//
