LOCUS       Exported                7231 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 7231)
  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..7231
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1196..1840
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
                     /note="/locus_tag=source:recombinant plasmid"
                     /note="This feature has 2 segments:
                        1: 1196 .. 1252 / #ffc0cb
                        2: 1253 .. 1840 / #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    506..509
                     /locus_tag="BfuAI site 1 overhang"
                     /label=PaqCI site 1 overhang
                     /label=misc_feature(12)
                     /note="color: #00ff00"
     misc_feature    514..520
                     /locus_tag="BfuA site 1"
                     /label=PaqCI site 1
                     /label=misc_feature(7)
                     /note="color: #00ff00"
     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..924
                     /locus_tag="BfuA site 2"
                     /label=PaqCI site 2
                     /label=misc_feature(5)
                     /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
                     /label=misc_feature(9)
                     /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
                     /label=misc_feature(2)
                     /note="color: #80ff80; direction: RIGHT"
     misc_feature    1016..1049
                     /locus_tag="lox2272"
                     /label=lox2272
                     /label=misc_feature(4)
                     /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: #8080ff"
     primer_bind     complement(1169..1203)
                     /label=2A_SA_R1
                     /note="color: black; sequence: 
                     ttcgtggcaactgtaaatgaatgagaaaaccggtt; added: 2021-02-21"
     primer_bind     1186..1213
                     /label=SA_2A_F1
                     /note="color: black; sequence: 
                     atttacagttgccacgaacttctctctg; added: 2021-01-08"
     CDS             1196..1252
                     /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     1234..1276
                     /label=KALTA4_F
                     /note="color: black; sequence: 
                     ggaagaaaaccccggtcctatgaaactgctctcatccatcgag; added: 
                     2021-01-29"
     primer_bind     complement(1235..1261)
                     /label=KAL_2A_R
                     /note="color: black; sequence: cagtttcataggaccggggttttcttc;
                     added: 2021-03-01"
     misc_feature    1253..1828
                     /label=KalTA4
                     /note="color: #00ffff"
     misc_feature    1253..1270
                     /label=KalTA4fw primer
                     /note="color: #ff9952"
     misc_feature    1712..1828
                     /label=TA4 (mit M to I)?
                     /note="color: #ff3d2e"
     misc_feature    1807..1828
                     /label=rev Primer
                     /note="color: #ff6ff1"
     primer_bind     complement(1820..1856)
                     /label=KALTA4_R
                     /note="color: black; sequence: 
                     ggctgagacttaattattagttacccgggaggatgtc; added: 2021-01-29"
     polyA_signal    1843..3018
                     /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"
     primer_bind     3019..3059
                     /label=Primer 1
                     /note="color: black; sequence: 
                     gaagcttgagatcctagtcaaagctttcaaccactccaggc; added: 
                     2021-10-01"
     misc_feature    3045..3409
                     /locus_tag="He1a_promoter"
                     /label=He1a_promoter
                     /note="color: #ff0000"
     primer_bind     complement(3425..3444)
                     /label=RFP NLS R
                     /note="color: black; sequence: cttcttctttggagccatgg; added:
                     2021-10-04"
     misc_signal     3427..3456
                     /locus_tag="NLS"
                     /label=NLS
                     /note="NLS"
                     /note="color: #ffc0cb"
     CDS             3457..4158
                     /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    4170..4768
                     /locus_tag="Danio b-Actin 3' UTR"
                     /label=Danio b-actin 3' UTR
                     /label=misc_feature(6)
                     /note="color: #0000ff"
     misc_feature    4749..5173
                     /locus_tag="Danio b-Actin poly(A) putative"
                     /label=Danio b-actin polyA
                     /label=misc_feature(8)
                     /note="color: #004080"
     misc_feature    complement(5180..5213)
                     /locus_tag="loxp(1)"
                     /label=loxp
                     /note="color: #00ff80; direction: LEFT"
     misc_feature    complement(5219..5252)
                     /locus_tag="lox2272(1)"
                     /label=lox2272
                     /note="color: #800080; direction: LEFT"
     misc_feature    complement(5259..5290)
                     /locus_tag="rox"
                     /label=rox
                     /label=misc_feature(1)
                     /note="color: #ff00ff; direction: LEFT"
     misc_feature    5300..5302
                     /locus_tag="TAL3(1)"
                     /label=TAL3
                     /label=TAL3(1)
                     /note="TAL3"
                     /note="color: #ffc0cb"
     misc_feature    5300..5302
                     /locus_tag="overhange3'5'"
                     /label=BspQI site 1 overhang
                     /label=misc_feature(10)
                     /note="color: #00ffff"
     misc_feature    5304..5310
                     /locus_tag="BspQ1 site1"
                     /label=misc_feature(15)
                     /note="color: #00ffff"
     intron          5319..5407
                     /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    5410..5416
                     /locus_tag="BspQ1site2"
                     /label=misc_feature(14)
                     /note="color: #00ffff"
     misc_feature    5418..5420
                     /locus_tag="BspQI site 2 overhang"
                     /label=BspQI site 2 overhang
                     /label=misc_feature(11)
                     /note="color: #00ffff"
     misc_feature    complement(5421..5440)
                     /locus_tag="Universal Guide 3'"
                     /label=misc_feature(13)
                     /note="color: #00ff00; direction: LEFT"
     misc_feature    5482..5696
                     /locus_tag="ITR(L)"
                     /label=ITR(L)
                     /note="ITR(L)"
                     /note="Tol2 ITR(L)"
                     /note="color: #80ffff"
     CDS             5761..6570
                     /locus_tag="Kan(R)"
                     /label=Kan(R)
                     /note="Kan(R)"
                     /note="color: #ffc0cb; direction: RIGHT"
     rep_origin      complement(6603..7231)
                     /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 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 cagttgccac
     1201 gaacttctct ctgttaaagc aagcaggaga cgtggaagaa aaccccggtc ctatgaaact
     1261 gctctcatcc atcgagcaag cctgcgacat ttgtcggctt aagaagctga aatgctccaa
     1321 ggaaaagccg aaatgtgcca aatgcctgaa gaacaattgg gaatgtcgtt actctcccaa
     1381 aaccaagcga agtccactca caagggctca tctgaccgaa gtggagagca ggctagagag
     1441 actggaacaa ctctttttgc tcatcttccc tagagaggac cttgacatga tcctcaagat
     1501 ggattctctc caggatatta aagccctttt gactggctta ttcgtccagg acaatgtgaa
     1561 caaagacgct gtgacagacc gattggcaag tgtcgagacc gatatgcctc tgacactgag
     1621 acagcacaga atcagcgcta cttcctcaag cgaagagtct tctaacaagg gacagagaca
     1681 gctgactgtt tcgagcaggt cgaccccgtc cccggccgac gccctggacg acggcgacct
     1741 ggacatgctg cctgctgatg ctctcgatga tttcgatctg gatatgctcc cggccgacgc
     1801 cctggacgac tacgacctgg acatcctccc gggtaactaa taattaagtc tcagccaccg
     1861 ttaactgaac atgtcaaaac ctgtggagac tgttgagatt tgatgttctg aaaagataaa
     1921 gcctataaat aaaatgttgc ccaaatttcc tgcctgatgt ttttctttgt ctttgctaca
     1981 tggctttgct gctcggatcg gctcactctg tgtatgccac gttcactttg tactctcctt
     2041 ctcacggtag gtttattatt tttagatgtg cagttagttt ctgtgaaata acacaccaca
     2101 cactgatatt gtctgtgcat tgacttggtg agtgcacatt gtttttgatc ttgacatatt
     2161 tatatttgat tgatcaggtg aactgtgtga atctaaagtg ctccatacag atgttctgca
     2221 ttgaaaatat tctcatttta ttagtggaag tgagtgtatg ccacatccaa tcaatttcag
     2281 caaacacccc agtatgattt aatgcaaaaa aatgaaggta tcaaacacgc attactactt
     2341 tgcagttaaa tatttaacat ttattccaac acgaaaaaaa gcagtaaata acactttgac
     2401 aaacacgtca ggacatctta tttttgtcac cctcacaggc aatttagtat aatatattat
     2461 atatatatat atatcatata ataatattca gtataatata tatatatata tcatattata
     2521 atattcagta taatataaaa cacaaacaca tatatgtata atataatata acatttttat
     2581 ttattgagat gcctctatgg accgtgttat aagaagtaaa gatcaggaga agtaaacatg
     2641 aagtgtaatt atgaatactg atgttaaatt aagctatgat gagttttcac tgttaattta
     2701 ccatctcaat taaatgttga tgcctccatg accaagttaa gcagatgaga ctgagacaac
     2761 tgtagaagac aagatgttca ctttgctgaa tatagctggc ttgacagtta tctatgactc
     2821 tataaatata tatatatttt tttttttata aaatgattta tttataacta tatatccatt
     2881 tctcagacag gtgcttcata tccctcactc ccgtagctgt ccatgctgga tctgtccccg
     2941 ttgtttttaa aaagctaaat aagttattaa catgactgca tccagcgagc caaacctgtc
     3001 tggtgtacag ctaccagaga agcttgagat cctagtcaaa gctttcaacc actccaggca
     3061 tagctagagc cactgatgca tctatagagc tgaagtaaag atagggttga tgcaaatcga
     3121 tttagaattt tcaatgtaga aaaaacaaaa taaaaagtct attcacttga tggatagatt
     3181 agtcaaacaa acatagagta ggcacttccc ttgactctgt aaacttttgg atttaacaga
     3241 ttgtgcactc cttttagtta aacacattag tccctgaaaa ctaagaccct aaccataatc
     3301 ccattgtaga cccacctact cgggtgaata taaagctcgc acagatcatc tcacagacat
     3361 cagaccttgg caaaggagca tcctgaacat ctctacacac tgaggcaacg aattcacctc
     3421 gagaccatgg ctccaaagaa gaagcgtaag gtaaccatga gcgagctgat taaggagaac
     3481 atgcacatga agctgtacat ggagggcacc gtggacaacc atcacttcaa gtgcacatcc
     3541 gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggt ggtcgagggc
     3601 ggccctctcc ccttcgcctt cgacatcctg gctactagct tcctctacgg cagcaagacc
     3661 ttcatcaacc acacccaggg catccccgac ttcttcaagc agtccttccc tgagggcttc
     3721 acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac ccaggacacc
     3781 agcctccagg acggctgcct catctacaac gtcaagatca gaggggtgaa cttcacatcc
     3841 aacggccctg tgatgcagaa gaaaacactc ggctgggagg ccttcaccga gacgctgtac
     3901 cccgctgacg gcggcctgga aggcagaaac gacatggccc tgaagctcgt gggcgggagc
     3961 catctgatcg caaacatcaa gaccacatat agatccaaga aacccgctaa gaacctcaag
     4021 atgcctggcg tctactatgt ggactacaga ctggaaagaa tcaaggaggc caacaacgag
     4081 acctacgtcg agcagcacga ggtggcagtg gccagatact gcgacctccc tagcaaactg
     4141 gggcacaagc tcaattaaag ctgagatcta cggactgtta ccacttcacg ccgactcaaa
     4201 ctgcgcagag aaacttcaaa cgacaacatt ggcatggctt ttgttatttt tggcgcttga
     4261 ctcaggatct aaaaactgga acggtgaagg tgacggcaat gttttttggc aaataagcat
     4321 ccccgaagtt ctacaatgca tctgaggact ccatgttttg ttttctttag tcattccaaa
     4381 tgtttgttaa atgcattgtt ccgaaactta tttgcctcta tgaaggctgc ccagtaattg
     4441 ggagcatact taacattgta gtattgtatg taaattatgt aacaaaacaa tgtctgggtt
     4501 tttgtacttt tcagccttaa aatcttgggt ttattttttt ttctttgttc caaaaaaacg
     4561 aagctttacc attcaagatg taaaggtttc cattccccct gggcatattg taaaagctgt
     4621 gtggaacgtg gcggtgccag acatttggtg gggccaacct gtacactgac taattcaatt
     4681 ccaataaaag tgcacatgta agacatccta cccctgtctc atttcctgtc ttggctctga
     4741 gtggcttcaa taaaaatggc ttttgtgaag ttgcaccatg ttaattacat ttcagcactt
     4801 tcacaccagg tttttgacta atttgaattt atcgaagttt gacctttttt tttttttttc
     4861 ttcaagctaa tttgaggtta tcaaaccatt tcattggagg aaacttaatg tttaaaagca
     4921 taaattcgaa acatgattca tttatttaac tgcttaatgg tgaaacaaac ttcaagtaat
     4981 acaccaagtg ttccctcata tgaaggtagc taactgcaaa cggccttaac tttccatctt
     5041 aagttgagct acataaaggc tttaacagga aggaactgcc agtttaaaag tcaaaccacc
     5101 atgactgata taaattcttg tccatataac cttgcgtttc acacaatata gctatctact
     5161 gctaaataaa ttagagctca taacttcgta taatgtatgc tatacgaagt tatgttagat
     5221 aacttcgtat aaagtatcct atacgaagtt atgcgagcta actttaaata attggcatta
     5281 tttaaagtta tgaatcgata agtgaagagc tatccggata tatatgtatg ttatcataac
     5341 tgtactggct atgtcaggtg gtaatgactg taacgttacg ttactcgttg taggcacgac
     5401 attgaatatg ctcttctccg ctgtggcccg aacgcctccc gctagcgatg ggatctaata
     5461 cgactcacta tagggagacc aagggaaaat agaatgaagt gatctccaaa aaataagtac
     5521 tttttgactg taaataaaat tgtaaggagt aaaaagtact tttttttcta aaaaaatgta
     5581 attaagtaaa agtaaaagta ttgattttta attgtactca agtaaagtaa aaatccccaa
     5641 aaataatact taagtacagt aatcaagtaa aattactcaa gtactttaca cctctgggat
     5701 ccagctgcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     5761 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
     5821 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
     5881 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     5941 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     6001 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     6061 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     6121 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     6181 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     6241 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     6301 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     6361 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     6421 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     6481 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     6541 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
     6601 caaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6661 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6721 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6781 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6841 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6901 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6961 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     7021 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     7081 aactgagata cctacagcgt gagctatgag aaagcgccac ggttcccgaa gggagaaagg
     7141 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7201 ggggaaacgc ctggtatctt tatagtcctg t
//
