LOCUS       Exported                7323 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 7323)
  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..7323
                     /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"
     source          3131..3518
                     /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    506..509
                     /locus_tag="BfuAI site 1 overhang"
                     /label=BfuAI site 1 overhang
                     /label=misc_feature(12)
                     /note="color: #00ff00"
     misc_feature    514..519
                     /locus_tag="BfuA site 1"
                     /label=BfuAI 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..923
                     /locus_tag="BfuA site 2"
                     /label=BfuAI 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"
     primer_bind     3111..3151
                     /label=Primer 1
                     /note="color: black; sequence: 
                     gaagcttgagatcctagtcaaagctttcaaccactccaggc; added: 
                     2021-10-01"
     misc_feature    3137..3501
                     /locus_tag="He1a_promoter"
                     /label=He1a_promoter
                     /note="color: #ff0000"
     primer_bind     complement(3517..3536)
                     /label=RFP NLS R
                     /note="color: black; sequence: cttcttctttggagccatgg; added:
                     2021-10-04"
     misc_signal     3519..3548
                     /locus_tag="NLS"
                     /label=NLS
                     /note="NLS"
                     /note="color: #ffc0cb"
     primer_bind     complement(3522..3545)
                     /label=HE1A-R
                     /note="color: black; sequence: taccttacgcttcttctttggagc; 
                     added: 2021-10-20"
     CDS             3549..4250
                     /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    4262..4860
                     /locus_tag="Danio b-Actin 3' UTR"
                     /label=Danio b-actin 3'UTR
                     /label=misc_feature(6)
                     /note="color: #0000ff"
     misc_feature    4841..5265
                     /locus_tag="Danio b-Actin poly(A) putative"
                     /label=Danio b-actin polyA
                     /label=misc_feature(8)
                     /note="color: #004080"
     misc_feature    complement(5272..5305)
                     /locus_tag="loxp(1)"
                     /label=loxp
                     /note="color: #00ff80; direction: LEFT"
     misc_feature    complement(5311..5344)
                     /locus_tag="lox2272(1)"
                     /label=lox2272
                     /note="color: #800080; direction: LEFT"
     misc_feature    complement(5351..5382)
                     /locus_tag="rox"
                     /label=rox
                     /label=misc_feature(1)
                     /note="color: #ff00ff; direction: LEFT"
     misc_feature    5392..5394
                     /locus_tag="TAL3(1)"
                     /label=TAL3
                     /label=TAL3(1)
                     /note="TAL3"
                     /note="color: #ffc0cb"
     misc_feature    5392..5394
                     /locus_tag="overhange3'5'"
                     /label=BspQ1 site 1 overhang
                     /label=misc_feature(10)
                     /note="color: #00ffff"
     misc_feature    5396..5402
                     /locus_tag="BspQ1 site1"
                     /label=misc_feature(15)
                     /note="color: #00ffff"
     intron          5411..5499
                     /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    5502..5508
                     /locus_tag="BspQ1site2"
                     /label=misc_feature(14)
                     /note="color: #00ffff"
     misc_feature    5510..5512
                     /locus_tag="BspQI site 2 overhang"
                     /label=BspQI site 2 overhang
                     /label=misc_feature(11)
                     /note="color: #00ffff"
     misc_feature    complement(5513..5532)
                     /locus_tag="Universal Guide 3'"
                     /label=3' Universal Guide
                     /label=misc_feature(13)
                     /note="color: #00ff00; direction: LEFT"
     misc_feature    5574..5788
                     /locus_tag="ITR(L)"
                     /label=ITR(L)
                     /note="ITR(L)"
                     /note="Tol2 ITR(L)"
                     /note="color: #80ffff"
     CDS             5853..6662
                     /locus_tag="Kan(R)"
                     /label=Kan(R)
                     /note="Kan(R)"
                     /note="color: #ffc0cb; direction: RIGHT"
     rep_origin      complement(6695..7323)
                     /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 aagctttcaa ccactccagg catagctaga gccactgatg catctataga
     3181 gctgaagtaa agatagggtt gatgcaaatc gatttagaat tttcaatgta gaaaaaacaa
     3241 aataaaaagt ctattcactt gatggataga ttagtcaaac aaacatagag taggcacttc
     3301 ccttgactct gtaaactttt ggatttaaca gattgtgcac tccttttagt taaacacatt
     3361 agtccctgaa aactaagacc ctaaccataa tcccattgta gacccaccta ctcgggtgaa
     3421 tataaagctc gcacagatca tctcacagac atcagacctt ggcaaaggag catcctgaac
     3481 atctctacac actgaggcaa cgaattcacc tcgagaccat ggctccaaag aagaagcgta
     3541 aggtaaccat gagcgagctg attaaggaga acatgcacat gaagctgtac atggagggca
     3601 ccgtggacaa ccatcacttc aagtgcacat ccgagggcga aggcaagccc tacgagggca
     3661 cccagaccat gagaatcaag gtggtcgagg gcggccctct ccccttcgcc ttcgacatcc
     3721 tggctactag cttcctctac ggcagcaaga ccttcatcaa ccacacccag ggcatccccg
     3781 acttcttcaa gcagtccttc cctgagggct tcacatggga gagagtcacc acatacgaag
     3841 acgggggcgt gctgaccgct acccaggaca ccagcctcca ggacggctgc ctcatctaca
     3901 acgtcaagat cagaggggtg aacttcacat ccaacggccc tgtgatgcag aagaaaacac
     3961 tcggctggga ggccttcacc gagacgctgt accccgctga cggcggcctg gaaggcagaa
     4021 acgacatggc cctgaagctc gtgggcggga gccatctgat cgcaaacatc aagaccacat
     4081 atagatccaa gaaacccgct aagaacctca agatgcctgg cgtctactat gtggactaca
     4141 gactggaaag aatcaaggag gccaacaacg agacctacgt cgagcagcac gaggtggcag
     4201 tggccagata ctgcgacctc cctagcaaac tggggcacaa gctcaattaa agctgagatc
     4261 tacggactgt taccacttca cgccgactca aactgcgcag agaaacttca aacgacaaca
     4321 ttggcatggc ttttgttatt tttggcgctt gactcaggat ctaaaaactg gaacggtgaa
     4381 ggtgacggca atgttttttg gcaaataagc atccccgaag ttctacaatg catctgagga
     4441 ctccatgttt tgttttcttt agtcattcca aatgtttgtt aaatgcattg ttccgaaact
     4501 tatttgcctc tatgaaggct gcccagtaat tgggagcata cttaacattg tagtattgta
     4561 tgtaaattat gtaacaaaac aatgtctggg tttttgtact tttcagcctt aaaatcttgg
     4621 gtttattttt ttttctttgt tccaaaaaaa cgaagcttta ccattcaaga tgtaaaggtt
     4681 tccattcccc ctgggcatat tgtaaaagct gtgtggaacg tggcggtgcc agacatttgg
     4741 tggggccaac ctgtacactg actaattcaa ttccaataaa agtgcacatg taagacatcc
     4801 tacccctgtc tcatttcctg tcttggctct gagtggcttc aataaaaatg gcttttgtga
     4861 agttgcacca tgttaattac atttcagcac tttcacacca ggtttttgac taatttgaat
     4921 ttatcgaagt ttgacctttt tttttttttt tcttcaagct aatttgaggt tatcaaacca
     4981 tttcattgga ggaaacttaa tgtttaaaag cataaattcg aaacatgatt catttattta
     5041 actgcttaat ggtgaaacaa acttcaagta atacaccaag tgttccctca tatgaaggta
     5101 gctaactgca aacggcctta actttccatc ttaagttgag ctacataaag gctttaacag
     5161 gaaggaactg ccagtttaaa agtcaaacca ccatgactga tataaattct tgtccatata
     5221 accttgcgtt tcacacaata tagctatcta ctgctaaata aattagagct cataacttcg
     5281 tataatgtat gctatacgaa gttatgttag ataacttcgt ataaagtatc ctatacgaag
     5341 ttatgcgagc taactttaaa taattggcat tatttaaagt tatgaatcga taagtgaaga
     5401 gctatccgga tatatatgta tgttatcata actgtactgg ctatgtcagg tggtaatgac
     5461 tgtaacgtta cgttactcgt tgtaggcacg acattgaata tgctcttctc cgctgtggcc
     5521 cgaacgcctc ccgctagcga tgggatctaa tacgactcac tatagggaga ccaagggaaa
     5581 atagaatgaa gtgatctcca aaaaataagt actttttgac tgtaaataaa attgtaagga
     5641 gtaaaaagta cttttttttc taaaaaaatg taattaagta aaagtaaaag tattgatttt
     5701 taattgtact caagtaaagt aaaaatcccc aaaaataata cttaagtaca gtaatcaagt
     5761 aaaattactc aagtacttta cacctctggg atccagctgc atcatgaaca ataaaactgt
     5821 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga
     5881 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata
     5941 atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt
     6001 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac
     6061 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg
     6121 atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag
     6181 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc
     6241 attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg
     6301 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg
     6361 gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt
     6421 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa
     6481 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc
     6541 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg
     6601 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct
     6661 aatcagaatt ggttaattgg ttgtaacact ggcaaaaagg atctaggtga agatcctttt
     6721 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6781 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6841 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6901 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     6961 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     7021 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     7081 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     7141 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     7201 agaaagcgcc acggttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     7261 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     7321 tgt
//
