LOCUS       Exported                7589 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 7589)
  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..7589
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1195..1251
                     /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
                     /label=misc_feature
                     /note="color: #00ffff"
     misc_feature    complement(506..509)
                     /locus_tag="BfuAI site 1 overhang"
                     /label=BfuAI site 1 overhang
                     /label=misc_feature(12)
                     /note="color: #00ff00; direction: LEFT"
     misc_feature    complement(514..519)
                     /locus_tag="BfuA site 1"
                     /label=misc_feature(7)
                     /note="color: #00ff00; direction: LEFT"
     promoter        520..641
                     /locus_tag="lacZ\promoter"
                     /label=lacZ promoter
                     /label=promoter
                     /note="/vntifkey=30"
                     /note="color: #ffc0cb"
     misc_feature    642..917
                     /locus_tag="lacZ'"
                     /label=lacZ
                     /label=misc_feature(3)
                     /note="/vntifkey=21"
                     /note="color: #ffc0cb"
     misc_feature    918..923
                     /locus_tag="BfuA site 2"
                     /label=misc_feature(5)
                     /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
                     /label=misc_feature(9)
                     /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
                     /label=misc_feature(2)
                     /note="color: #80ff80; direction: RIGHT"
     misc_feature    1015..1048
                     /locus_tag="lox2272"
                     /label=lox2272
                     /label=misc_feature(4)
                     /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: #8080ff"
     primer_bind     complement(1168..1202)
                     /label=2A_SA_R1
                     /note="color: black; sequence: 
                     ttcgtggcaactgtaaatgaatgagaaaaccggtt; added: 2021-02-21"
     primer_bind     1185..1212
                     /label=SA_2A_F1
                     /note="color: black; sequence: 
                     atttacagttgccacgaacttctctctg; added: 2021-01-08"
     CDS             1195..1251
                     /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(1234..1260)
                     /label=RFP_2A_R
                     /note="color: black; sequence: ggaggaggcaggaccggggttttcttc;
                     added: 2021-01-08"
     primer_bind     1243..1269
                     /label=2A_RFP_F
                     /note="color: black; sequence: cccggtcctgcctcctccgaggacgtc;
                     added: 2021-01-08"
     CDS             1252..1926
                     /locus_tag="mRFP*"
                     /label=mRFP*
                     /note="mRFP*"
                     /note="color: #ff0000; direction: RIGHT"
     primer_bind     complement(1567..1604)
                     /label=mRFP_R
                     /note="color: black; sequence: 
                     atgaactcgccgtcctgcagggaggagtcctgggtcac; added: 2021-03-01"
     polyA_signal    1935..3110
                     /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: #ff8000"
     misc_feature    3154..3514
                     /locus_tag="gamma-crystallin"
                     /label=gamma-crystallin
                     /note="gamma-crystallin"
                     /note="Xenopus"
                     /note="color: #ffc0cb"
     intron          3567..3764
                     /locus_tag="mIntron"
                     /label=mIntron
                     /note="mIntron"
                     /note="carp bactin shortened intron I"
                     /note="color: #ffc0cb"
     primer_bind     complement(3735..3761)
                     /label=2A_SA_R1
                     /note="color: black; sequence: 
                     ttcgtggcaactgtaaatgaatgagaaaaccggtt; added: 2021-02-21"
     misc_signal     3785..3814
                     /locus_tag="NLS"
                     /label=NLS
                     /note="NLS"
                     /note="color: #ffc0cb"
     CDS             3815..4516
                     /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    4528..5126
                     /locus_tag="Danio b-Actin 3' UTR"
                     /label=Danio b-actin 3'UTR
                     /label=misc_feature(6)
                     /note="color: #0000ff"
     misc_feature    5107..5531
                     /locus_tag="Danio b-Actin poly(A) putative"
                     /label=Danio b-actin polyA
                     /label=misc_feature(8)
                     /note="color: #004080"
     misc_feature    complement(5538..5571)
                     /locus_tag="loxp(1)"
                     /label=loxp
                     /note="color: #00ff80; direction: LEFT"
     misc_feature    complement(5577..5610)
                     /locus_tag="lox2272(1)"
                     /label=lox2272
                     /note="color: #800080; direction: LEFT"
     misc_feature    complement(5617..5648)
                     /locus_tag="rox"
                     /label=rox
                     /label=misc_feature(1)
                     /note="color: #ff00ff; direction: LEFT"
     misc_feature    5658..5660
                     /locus_tag="TAL3(1)"
                     /label=TAL3
                     /label=TAL3(1)
                     /note="TAL3"
                     /note="color: #ffc0cb"
     misc_feature    5658..5660
                     /locus_tag="overhange3'5'"
                     /label=BspQ1 site 1 overhang
                     /label=misc_feature(10)
                     /note="color: #00ffff"
     misc_feature    5662..5668
                     /locus_tag="BspQ1 site1"
                     /label=misc_feature(15)
                     /note="color: #00ffff"
     intron          5677..5765
                     /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    5768..5774
                     /locus_tag="BspQ1site2"
                     /label=misc_feature(14)
                     /note="color: #00ffff"
     misc_feature    5776..5778
                     /locus_tag="BspQI site 2 overhang"
                     /label=BspQI site 2 overhang
                     /label=misc_feature(11)
                     /note="color: #00ffff"
     misc_feature    complement(5779..5798)
                     /locus_tag="Universal Guide 3'"
                     /label=3' Universal Guide
                     /label=misc_feature(13)
                     /note="color: #00ff00; direction: LEFT"
     misc_feature    5840..6054
                     /locus_tag="ITR(L)"
                     /label=ITR(L)
                     /note="ITR(L)"
                     /note="Tol2 ITR(L)"
                     /note="color: #80ffff"
     CDS             6119..6928
                     /locus_tag="Kan(R)"
                     /label=Kan(R)
                     /note="Kan(R)"
                     /note="color: #ffc0cb; direction: RIGHT"
     rep_origin      complement(6961..7589)
                     /direction=LEFT
                     /locus_tag="ColE1 Ori(1)"
                     /label=ColE1 Ori
                     /label=ColE1 Ori(1)
                     /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 agttgccacg
     1201 aacttctctc tgttaaagca agcaggagac gtggaagaaa accccggtcc tgcctcctcc
     1261 gaggacgtca tcaaggagtt catgcgcttc aaggtgcgca tggagggctc cgtgaacggc
     1321 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     1381 aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct gtcccctcag
     1441 ttccagtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
     1501 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
     1561 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg
     1621 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     1681 gcctccaccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagatgagg
     1741 ctgaagctga aggacggcgg ccactacgac gccgaggtca agaccaccta catggccaag
     1801 aagcccgtgc agctgcccgg cgcctacaag accgacatca agctggacat cacctcccac
     1861 aacgaggact acaccatcgt ggaacagtac gagcgcgccg agggccgcca ctccaccgga
     1921 gcctaagact agtaattaag tctcagccac cgttaactga acatgtcaaa acctgtggag
     1981 actgttgaga tttgatgttc tgaaaagata aagcctataa ataaaatgtt gcccaaattt
     2041 cctgcctgat gtttttcttt gtctttgcta catggctttg ctgctcggat cggctcactc
     2101 tgtgtatgcc acgttcactt tgtactctcc ttctcacggt aggtttatta tttttagatg
     2161 tgcagttagt ttctgtgaaa taacacacca cacactgata ttgtctgtgc attgacttgg
     2221 tgagtgcaca ttgtttttga tcttgacata tttatatttg attgatcagg tgaactgtgt
     2281 gaatctaaag tgctccatac agatgttctg cattgaaaat attctcattt tattagtgga
     2341 agtgagtgta tgccacatcc aatcaatttc agcaaacacc ccagtatgat ttaatgcaaa
     2401 aaaatgaagg tatcaaacac gcattactac tttgcagtta aatatttaac atttattcca
     2461 acacgaaaaa aagcagtaaa taacactttg acaaacacgt caggacatct tatttttgtc
     2521 accctcacag gcaatttagt ataatatatt atatatatat atatatcata taataatatt
     2581 cagtataata tatatatata tatcatatta taatattcag tataatataa aacacaaaca
     2641 catatatgta taatataata taacattttt atttattgag atgcctctat ggaccgtgtt
     2701 ataagaagta aagatcagga gaagtaaaca tgaagtgtaa ttatgaatac tgatgttaaa
     2761 ttaagctatg atgagttttc actgttaatt taccatctca attaaatgtt gatgcctcca
     2821 tgaccaagtt aagcagatga gactgagaca actgtagaag acaagatgtt cactttgctg
     2881 aatatagctg gcttgacagt tatctatgac tctataaata tatatatatt ttttttttta
     2941 taaaatgatt tatttataac tatatatcca tttctcagac aggtgcttca tatccctcac
     3001 tcccgtagct gtccatgctg gatctgtccc cgttgttttt aaaaagctaa ataagttatt
     3061 aacatgactg catccagcga gccaaacctg tctggtgtac agctaccaga gaagcttgag
     3121 atcctagtca ccggcgttcc tagttcttta aactgttaca aagtgtttgg accagcaact
     3181 gaattgcact gtaaattacc acatcagggt ctccggggtc aacgctgcta agcaatggac
     3241 tggaaaattt gtgacattga gtttttggaa taactgggaa tgctggataa ctctttgggt
     3301 aaagctatgg aacaaaagta ccagcatgaa ctaagaaatg actgtcacag cacaacatca
     3361 aacacaatgc aagacaaagg ttctctgctt ttgccttact tgacagaaaa agtaagctct
     3421 aggttcatcg catgcagaca gcagcaatac atgacatata taaaggctgg ctcttttctc
     3481 ccttccattg aactgaaact tccactcagt cagacttgcg ggggatccac agcctgacta
     3541 actcgacaaa cccccccaaa cctaaggtga gttgatcttt aagcttttta cattttcagc
     3601 tcgcatatat caattcgcgc gcattgtcac actagcaccc actagcggtc agactgcaga
     3661 caaacaggaa gctgactcca catggtcaca tgctcactga agtgttgact tccctgacag
     3721 ctgtgcactt tctaaaccgg ttttctcatt catttacagt tcagccggaa ttcacctcga
     3781 gaccatggct ccaaagaaga agcgtaaggt aaccatgagc gagctgatta aggagaacat
     3841 gcacatgaag ctgtacatgg agggcaccgt ggacaaccat cacttcaagt gcacatccga
     3901 gggcgaaggc aagccctacg agggcaccca gaccatgaga atcaaggtgg tcgagggcgg
     3961 ccctctcccc ttcgccttcg acatcctggc tactagcttc ctctacggca gcaagacctt
     4021 catcaaccac acccagggca tccccgactt cttcaagcag tccttccctg agggcttcac
     4081 atgggagaga gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag
     4141 cctccaggac ggctgcctca tctacaacgt caagatcaga ggggtgaact tcacatccaa
     4201 cggccctgtg atgcagaaga aaacactcgg ctgggaggcc ttcaccgaga cgctgtaccc
     4261 cgctgacggc ggcctggaag gcagaaacga catggccctg aagctcgtgg gcgggagcca
     4321 tctgatcgca aacatcaaga ccacatatag atccaagaaa cccgctaaga acctcaagat
     4381 gcctggcgtc tactatgtgg actacagact ggaaagaatc aaggaggcca acaacgagac
     4441 ctacgtcgag cagcacgagg tggcagtggc cagatactgc gacctcccta gcaaactggg
     4501 gcacaagctc aattaaagct gagatctacg gactgttacc acttcacgcc gactcaaact
     4561 gcgcagagaa acttcaaacg acaacattgg catggctttt gttatttttg gcgcttgact
     4621 caggatctaa aaactggaac ggtgaaggtg acggcaatgt tttttggcaa ataagcatcc
     4681 ccgaagttct acaatgcatc tgaggactcc atgttttgtt ttctttagtc attccaaatg
     4741 tttgttaaat gcattgttcc gaaacttatt tgcctctatg aaggctgccc agtaattggg
     4801 agcatactta acattgtagt attgtatgta aattatgtaa caaaacaatg tctgggtttt
     4861 tgtacttttc agccttaaaa tcttgggttt attttttttt ctttgttcca aaaaaacgaa
     4921 gctttaccat tcaagatgta aaggtttcca ttccccctgg gcatattgta aaagctgtgt
     4981 ggaacgtggc ggtgccagac atttggtggg gccaacctgt acactgacta attcaattcc
     5041 aataaaagtg cacatgtaag acatcctacc cctgtctcat ttcctgtctt ggctctgagt
     5101 ggcttcaata aaaatggctt ttgtgaagtt gcaccatgtt aattacattt cagcactttc
     5161 acaccaggtt tttgactaat ttgaatttat cgaagtttga cctttttttt tttttttctt
     5221 caagctaatt tgaggttatc aaaccatttc attggaggaa acttaatgtt taaaagcata
     5281 aattcgaaac atgattcatt tatttaactg cttaatggtg aaacaaactt caagtaatac
     5341 accaagtgtt ccctcatatg aaggtagcta actgcaaacg gccttaactt tccatcttaa
     5401 gttgagctac ataaaggctt taacaggaag gaactgccag tttaaaagtc aaaccaccat
     5461 gactgatata aattcttgtc catataacct tgcgtttcac acaatatagc tatctactgc
     5521 taaataaatt agagctcata acttcgtata atgtatgcta tacgaagtta tgttagataa
     5581 cttcgtataa agtatcctat acgaagttat gcgagctaac tttaaataat tggcattatt
     5641 taaagttatg aatcgataag tgaagagcta tccggatata tatgtatgtt atcataactg
     5701 tactggctat gtcaggtggt aatgactgta acgttacgtt actcgttgta ggcacgacat
     5761 tgaatatgct cttctccgct gtggcccgaa cgcctcccgc tagcgatggg atctaatacg
     5821 actcactata gggagaccaa gggaaaatag aatgaagtga tctccaaaaa ataagtactt
     5881 tttgactgta aataaaattg taaggagtaa aaagtacttt tttttctaaa aaaatgtaat
     5941 taagtaaaag taaaagtatt gatttttaat tgtactcaag taaagtaaaa atccccaaaa
     6001 ataatactta agtacagtaa tcaagtaaaa ttactcaagt actttacacc tctgggatcc
     6061 agctgcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     6121 gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt
     6181 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt
     6241 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     6301 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     6361 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     6421 aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     6481 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     6541 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     6601 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     6661 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     6721 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     6781 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     6841 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     6901 tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca
     6961 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     7021 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     7081 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     7141 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     7201 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     7261 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     7321 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     7381 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     7441 ctgagatacc tacagcgtga gctatgagaa agcgccacgg ttcccgaagg gagaaaggcg
     7501 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7561 ggaaacgcct ggtatcttta tagtcctgt
//
