LOCUS       Exported                7229 bp ds-DNA     circular SYN 09-NOV-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7229)
  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..7229
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1050..4167
                     /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=PaqCI site 1 overhang
                     /note="color: #00ff00"
     misc_feature    514..520
                     /locus_tag="BfuA site 1"
                     /label=PaqCI 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..924
                     /locus_tag="BfuA site 2"
                     /label=PaqCI site 2
                     /note="color: #00ffff"
     misc_feature    complement(929..939)
                     /locus_tag="TAL3"
                     /label=TAL3
                     /note="TAL3"
                     /note="color: #ffc0cb; direction: LEFT"
     misc_feature    929..932
                     /locus_tag="overhang5'3'"
                     /label=PaqCI site 2 overhang
                     /note="color: #00ffff"
     misc_feature    941..972
                     /locus_tag="rox(1)"
                     /label=rox
                     /note="color: #ff0080; direction: RIGHT"
     misc_feature    978..1011
                     /locus_tag="loxp"
                     /label=loxp
                     /note="color: #80ff80; direction: RIGHT"
     misc_feature    1016..1049
                     /locus_tag="lox2272"
                     /label=lox2272
                     /note="color: #8080ff; direction: RIGHT"
     intron          1061..1193
                     /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(1169..1203)
                     /label=2A_SA_R0
                     /note="color: black; sequence: 
                     agttcgtggcctgtaaatgaatgagaaaaccggtt; added: 2021-03-01"
     primer_bind     1184..1211
                     /label=SA_2A_F0
                     /note="color: black; sequence: 
                     tcatttacaggccacgaacttctctctg; added: 2021-03-01"
     CDS             1194..1250
                     /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     1232..1274
                     /label=KALTA4_F
                     /note="color: black; sequence: 
                     ggaagaaaaccccggtcctatgaaactgctctcatccatcgag; added: 
                     2021-01-29"
     primer_bind     complement(1233..1259)
                     /label=KAL_2A_R
                     /note="color: black; sequence: cagtttcataggaccggggttttcttc;
                     added: 2021-03-01"
     misc_feature    1251..1826
                     /label=KalTA4
                     /note="color: #00ffff"
     misc_feature    1251..1268
                     /label=KalTA4fw primer
                     /note="color: #ff9952"
     misc_feature    1710..1826
                     /label=TA4 (mit M to I)?
                     /note="color: #ff3d2e"
     misc_feature    1805..1826
                     /label=rev Primer
                     /note="color: #ff6ff1"
     primer_bind     complement(1818..1854)
                     /label=KALTA4_R
                     /note="color: black; sequence: 
                     ggctgagacttaattattagttacccgggaggatgtc; added: 2021-01-29"
     polyA_signal    1841..3016
                     /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     3017..3057
                     /label=Primer 1
                     /note="color: black; sequence: 
                     gaagcttgagatcctagtcaaagctttcaaccactccaggc; added: 
                     2021-10-01"
     misc_feature    3043..3407
                     /locus_tag="He1a_promoter"
                     /label=He1a_promoter
                     /note="color: #ff0000"
     primer_bind     complement(3423..3442)
                     /label=RFP NLS R
                     /note="color: black; sequence: cttcttctttggagccatgg; added:
                     2021-10-04"
     misc_signal     3425..3454
                     /locus_tag="NLS"
                     /label=NLS
                     /note="NLS"
                     /note="color: #ffc0cb"
     CDS             3455..4156
                     /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    4168..4766
                     /locus_tag="Danio b-Actin 3' UTR"
                     /label=Danio b-Actin 3' UTR
                     /note="color: #0000ff"
     misc_feature    4747..5171
                     /locus_tag="Danio b-Actin poly(A) putative"
                     /label=Danio b-Actin poly(A) putative
                     /note="color: #004080"
     misc_feature    complement(5178..5211)
                     /locus_tag="loxp(1)"
                     /label=loxp
                     /note="color: #00ff80; direction: LEFT"
     misc_feature    complement(5217..5250)
                     /locus_tag="lox2272(1)"
                     /label=lox2272
                     /note="color: #800080; direction: LEFT"
     misc_feature    complement(5257..5288)
                     /locus_tag="rox"
                     /label=rox
                     /note="color: #ff00ff; direction: LEFT"
     misc_feature    5298..5300
                     /locus_tag="TAL3(1)"
                     /label=TAL3
                     /note="TAL3"
                     /note="color: #ffc0cb"
     misc_feature    5298..5300
                     /locus_tag="overhange3'5'"
                     /label=BspQ1 site 1 overhang 3'5'
                     /note="color: #00ffff"
     misc_feature    5302..5308
                     /locus_tag="BspQ1 site1"
                     /label=BspQ1 site1
                     /note="color: #00ffff"
     intron          5317..5405
                     /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    5408..5414
                     /locus_tag="BspQ1site2"
                     /label=BspQ1site2
                     /note="color: #00ffff"
     misc_feature    complement(5416..5438)
                     /locus_tag="Universal Guide 3'"
                     /label=Universal Guide 3'
                     /note="color: #00ff00; direction: LEFT"
     misc_feature    5416..5418
                     /locus_tag="BspQI site 2 overhang"
                     /label=BspQI site 2 overhang
                     /note="color: #00ffff"
     misc_feature    5480..5694
                     /locus_tag="ITR(L)"
                     /label=ITR(L)
                     /note="ITR(L)"
                     /note="Tol2 ITR(L)"
                     /note="color: #80ffff"
     CDS             5759..6568
                     /locus_tag="Kan(R)"
                     /label=Kan(R)
                     /note="Kan(R)"
                     /note="color: #ffc0cb; direction: RIGHT"
     rep_origin      complement(6601..7229)
                     /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 cgaatggcac ctgccacact tccggccgca taactttaaa taatgccaat
      961 tatttaaagt tactatcata acttcgtata gcatacatta tacgaagtta tcaccataac
     1021 ttcgtatagg atactttata cgaagttatc ctaggcttac cacccactag cggtcagact
     1081 gcagattgca gcacgaaaca ggaagctgac tccacatggt cacatgctca ctgaagtgtt
     1141 gacttccctg acagctgtgc actttctaaa ccggttttct cattcattta caggccacga
     1201 acttctctct gttaaagcaa gcaggagacg tggaagaaaa ccccggtcct atgaaactgc
     1261 tctcatccat cgagcaagcc tgcgacattt gtcggcttaa gaagctgaaa tgctccaagg
     1321 aaaagccgaa atgtgccaaa tgcctgaaga acaattggga atgtcgttac tctcccaaaa
     1381 ccaagcgaag tccactcaca agggctcatc tgaccgaagt ggagagcagg ctagagagac
     1441 tggaacaact ctttttgctc atcttcccta gagaggacct tgacatgatc ctcaagatgg
     1501 attctctcca ggatattaaa gcccttttga ctggcttatt cgtccaggac aatgtgaaca
     1561 aagacgctgt gacagaccga ttggcaagtg tcgagaccga tatgcctctg acactgagac
     1621 agcacagaat cagcgctact tcctcaagcg aagagtcttc taacaaggga cagagacagc
     1681 tgactgtttc gagcaggtcg accccgtccc cggccgacgc cctggacgac ggcgacctgg
     1741 acatgctgcc tgctgatgct ctcgatgatt tcgatctgga tatgctcccg gccgacgccc
     1801 tggacgacta cgacctggac atcctcccgg gtaactaata attaagtctc agccaccgtt
     1861 aactgaacat gtcaaaacct gtggagactg ttgagatttg atgttctgaa aagataaagc
     1921 ctataaataa aatgttgccc aaatttcctg cctgatgttt ttctttgtct ttgctacatg
     1981 gctttgctgc tcggatcggc tcactctgtg tatgccacgt tcactttgta ctctccttct
     2041 cacggtaggt ttattatttt tagatgtgca gttagtttct gtgaaataac acaccacaca
     2101 ctgatattgt ctgtgcattg acttggtgag tgcacattgt ttttgatctt gacatattta
     2161 tatttgattg atcaggtgaa ctgtgtgaat ctaaagtgct ccatacagat gttctgcatt
     2221 gaaaatattc tcattttatt agtggaagtg agtgtatgcc acatccaatc aatttcagca
     2281 aacaccccag tatgatttaa tgcaaaaaaa tgaaggtatc aaacacgcat tactactttg
     2341 cagttaaata tttaacattt attccaacac gaaaaaaagc agtaaataac actttgacaa
     2401 acacgtcagg acatcttatt tttgtcaccc tcacaggcaa tttagtataa tatattatat
     2461 atatatatat atcatataat aatattcagt ataatatata tatatatatc atattataat
     2521 attcagtata atataaaaca caaacacata tatgtataat ataatataac atttttattt
     2581 attgagatgc ctctatggac cgtgttataa gaagtaaaga tcaggagaag taaacatgaa
     2641 gtgtaattat gaatactgat gttaaattaa gctatgatga gttttcactg ttaatttacc
     2701 atctcaatta aatgttgatg cctccatgac caagttaagc agatgagact gagacaactg
     2761 tagaagacaa gatgttcact ttgctgaata tagctggctt gacagttatc tatgactcta
     2821 taaatatata tatatttttt tttttataaa atgatttatt tataactata tatccatttc
     2881 tcagacaggt gcttcatatc cctcactccc gtagctgtcc atgctggatc tgtccccgtt
     2941 gtttttaaaa agctaaataa gttattaaca tgactgcatc cagcgagcca aacctgtctg
     3001 gtgtacagct accagagaag cttgagatcc tagtcaaagc tttcaaccac tccaggcata
     3061 gctagagcca ctgatgcatc tatagagctg aagtaaagat agggttgatg caaatcgatt
     3121 tagaattttc aatgtagaaa aaacaaaata aaaagtctat tcacttgatg gatagattag
     3181 tcaaacaaac atagagtagg cacttccctt gactctgtaa acttttggat ttaacagatt
     3241 gtgcactcct tttagttaaa cacattagtc cctgaaaact aagaccctaa ccataatccc
     3301 attgtagacc cacctactcg ggtgaatata aagctcgcac agatcatctc acagacatca
     3361 gaccttggca aaggagcatc ctgaacatct ctacacactg aggcaacgaa ttcacctcga
     3421 gaccatggct ccaaagaaga agcgtaaggt aaccatgagc gagctgatta aggagaacat
     3481 gcacatgaag ctgtacatgg agggcaccgt ggacaaccat cacttcaagt gcacatccga
     3541 gggcgaaggc aagccctacg agggcaccca gaccatgaga atcaaggtgg tcgagggcgg
     3601 ccctctcccc ttcgccttcg acatcctggc tactagcttc ctctacggca gcaagacctt
     3661 catcaaccac acccagggca tccccgactt cttcaagcag tccttccctg agggcttcac
     3721 atgggagaga gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag
     3781 cctccaggac ggctgcctca tctacaacgt caagatcaga ggggtgaact tcacatccaa
     3841 cggccctgtg atgcagaaga aaacactcgg ctgggaggcc ttcaccgaga cgctgtaccc
     3901 cgctgacggc ggcctggaag gcagaaacga catggccctg aagctcgtgg gcgggagcca
     3961 tctgatcgca aacatcaaga ccacatatag atccaagaaa cccgctaaga acctcaagat
     4021 gcctggcgtc tactatgtgg actacagact ggaaagaatc aaggaggcca acaacgagac
     4081 ctacgtcgag cagcacgagg tggcagtggc cagatactgc gacctcccta gcaaactggg
     4141 gcacaagctc aattaaagct gagatctacg gactgttacc acttcacgcc gactcaaact
     4201 gcgcagagaa acttcaaacg acaacattgg catggctttt gttatttttg gcgcttgact
     4261 caggatctaa aaactggaac ggtgaaggtg acggcaatgt tttttggcaa ataagcatcc
     4321 ccgaagttct acaatgcatc tgaggactcc atgttttgtt ttctttagtc attccaaatg
     4381 tttgttaaat gcattgttcc gaaacttatt tgcctctatg aaggctgccc agtaattggg
     4441 agcatactta acattgtagt attgtatgta aattatgtaa caaaacaatg tctgggtttt
     4501 tgtacttttc agccttaaaa tcttgggttt attttttttt ctttgttcca aaaaaacgaa
     4561 gctttaccat tcaagatgta aaggtttcca ttccccctgg gcatattgta aaagctgtgt
     4621 ggaacgtggc ggtgccagac atttggtggg gccaacctgt acactgacta attcaattcc
     4681 aataaaagtg cacatgtaag acatcctacc cctgtctcat ttcctgtctt ggctctgagt
     4741 ggcttcaata aaaatggctt ttgtgaagtt gcaccatgtt aattacattt cagcactttc
     4801 acaccaggtt tttgactaat ttgaatttat cgaagtttga cctttttttt tttttttctt
     4861 caagctaatt tgaggttatc aaaccatttc attggaggaa acttaatgtt taaaagcata
     4921 aattcgaaac atgattcatt tatttaactg cttaatggtg aaacaaactt caagtaatac
     4981 accaagtgtt ccctcatatg aaggtagcta actgcaaacg gccttaactt tccatcttaa
     5041 gttgagctac ataaaggctt taacaggaag gaactgccag tttaaaagtc aaaccaccat
     5101 gactgatata aattcttgtc catataacct tgcgtttcac acaatatagc tatctactgc
     5161 taaataaatt agagctcata acttcgtata atgtatgcta tacgaagtta tgttagataa
     5221 cttcgtataa agtatcctat acgaagttat gcgagctaac tttaaataat tggcattatt
     5281 taaagttatg aatcgataag tgaagagcta tccggatata tatgtatgtt atcataactg
     5341 tactggctat gtcaggtggt aatgactgta acgttacgtt actcgttgta ggcacgacat
     5401 tgaatatgct cttctccgct gtggcccgaa cgcctcccgc tagcgatggg atctaatacg
     5461 actcactata gggagaccaa gggaaaatag aatgaagtga tctccaaaaa ataagtactt
     5521 tttgactgta aataaaattg taaggagtaa aaagtacttt tttttctaaa aaaatgtaat
     5581 taagtaaaag taaaagtatt gatttttaat tgtactcaag taaagtaaaa atccccaaaa
     5641 ataatactta agtacagtaa tcaagtaaaa ttactcaagt actttacacc tctgggatcc
     5701 agctgcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     5761 gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt
     5821 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt
     5881 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     5941 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     6001 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     6061 aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     6121 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     6181 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     6241 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     6301 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     6361 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     6421 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     6481 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     6541 tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca
     6601 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6661 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6721 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6781 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6841 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6901 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6961 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     7021 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     7081 ctgagatacc tacagcgtga gctatgagaa agcgccacgg ttcccgaagg gagaaaggcg
     7141 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7201 ggaaacgcct ggtatcttta tagtcctgt
//
