LOCUS       Exported                7230 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 7230)
  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..7230
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1050..4168
                     /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"
     primer_bind     630..647
                     /label=R1
                     /note="color: black; sequence: CAggAAACAgCTATgACC; added: 
                     2021-10-11"
     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..1202)
                     /label=2A_SA_R2
                     /note="color: black; sequence: 
                     ttcgtggcgctgtaaatgaatgagaaaaccggtt; added: 2021-03-01"
     primer_bind     1186..1212
                     /label=SA_2A_F2
                     /note="color: black; sequence: atttacagcgccacgaacttctctctg;
                     added: 2021-03-01"
     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     1233..1275
                     /label=KALTA4_F
                     /note="color: black; sequence: 
                     ggaagaaaaccccggtcctatgaaactgctctcatccatcgag; added: 
                     2021-01-29"
     primer_bind     complement(1234..1260)
                     /label=KAL_2A_R
                     /note="color: black; sequence: cagtttcataggaccggggttttcttc;
                     added: 2021-03-01"
     misc_feature    1252..1827
                     /label=KalTA4
                     /note="color: #00ffff"
     misc_feature    1252..1269
                     /label=KalTA4fw primer
                     /note="color: #ff9952"
     misc_feature    1711..1827
                     /label=TA4 (mit M to I)?
                     /note="color: #ff3d2e"
     misc_feature    1806..1827
                     /label=rev Primer
                     /note="color: #ff6ff1"
     primer_bind     complement(1819..1855)
                     /label=KALTA4_R
                     /note="color: black; sequence: 
                     ggctgagacttaattattagttacccgggaggatgtc; added: 2021-01-29"
     polyA_signal    1842..3017
                     /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     3018..3058
                     /label=Primer 1
                     /note="color: black; sequence: 
                     gaagcttgagatcctagtcaaagctttcaaccactccaggc; added: 
                     2021-10-01"
     misc_feature    3044..3408
                     /locus_tag="He1a_promoter"
                     /label=He1a_promoter
                     /note="color: #ff0000"
     primer_bind     complement(3424..3443)
                     /label=RFP NLS R
                     /note="color: black; sequence: cttcttctttggagccatgg; added:
                     2021-10-04"
     misc_signal     3426..3455
                     /locus_tag="NLS"
                     /label=NLS
                     /note="NLS"
                     /note="color: #ffc0cb"
     CDS             3456..4157
                     /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    4169..4767
                     /locus_tag="Danio b-Actin 3' UTR"
                     /label=Danio b-Actin 3' UTR
                     /note="color: #0000ff"
     misc_feature    4748..5172
                     /locus_tag="Danio b-Actin poly(A) putative"
                     /label=Danio b-Actin poly(A) putative
                     /note="color: #004080"
     misc_feature    complement(5179..5212)
                     /locus_tag="loxp(1)"
                     /label=loxp
                     /note="color: #00ff80; direction: LEFT"
     misc_feature    complement(5218..5251)
                     /locus_tag="lox2272(1)"
                     /label=lox2272
                     /note="color: #800080; direction: LEFT"
     misc_feature    complement(5258..5289)
                     /locus_tag="rox"
                     /label=rox
                     /note="color: #ff00ff; direction: LEFT"
     misc_feature    5299..5301
                     /locus_tag="TAL3(1)"
                     /label=TAL3
                     /note="TAL3"
                     /note="color: #ffc0cb"
     misc_feature    5299..5301
                     /locus_tag="overhange3'5'"
                     /label=BspQI site 1 overhang 3'5'
                     /note="color: #00ffff"
     misc_feature    5303..5309
                     /locus_tag="BspQ1 site1"
                     /label=BspQ1 site1
                     /note="color: #00ffff"
     intron          5318..5406
                     /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    5409..5415
                     /locus_tag="BspQ1site2"
                     /label=BspQ1site2
                     /note="color: #00ffff"
     misc_feature    5417..5419
                     /locus_tag="BspQI site 2 overhang"
                     /label=BspQI site 2 overhang
                     /note="color: #00ffff"
     misc_feature    complement(5420..5439)
                     /locus_tag="Universal Guide 3'"
                     /label=Universal Guide 3'
                     /note="color: #00ff00; direction: LEFT"
     misc_feature    5481..5695
                     /locus_tag="ITR(L)"
                     /label=ITR(L)
                     /note="ITR(L)"
                     /note="Tol2 ITR(L)"
                     /note="color: #80ffff"
     CDS             5760..6569
                     /locus_tag="Kan(R)"
                     /label=Kan(R)
                     /note="Kan(R)"
                     /note="color: #ffc0cb; direction: RIGHT"
     rep_origin      complement(6602..7230)
                     /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 cagcgccacg
     1201 aacttctctc tgttaaagca agcaggagac gtggaagaaa accccggtcc tatgaaactg
     1261 ctctcatcca tcgagcaagc ctgcgacatt tgtcggctta agaagctgaa atgctccaag
     1321 gaaaagccga aatgtgccaa atgcctgaag aacaattggg aatgtcgtta ctctcccaaa
     1381 accaagcgaa gtccactcac aagggctcat ctgaccgaag tggagagcag gctagagaga
     1441 ctggaacaac tctttttgct catcttccct agagaggacc ttgacatgat cctcaagatg
     1501 gattctctcc aggatattaa agcccttttg actggcttat tcgtccagga caatgtgaac
     1561 aaagacgctg tgacagaccg attggcaagt gtcgagaccg atatgcctct gacactgaga
     1621 cagcacagaa tcagcgctac ttcctcaagc gaagagtctt ctaacaaggg acagagacag
     1681 ctgactgttt cgagcaggtc gaccccgtcc ccggccgacg ccctggacga cggcgacctg
     1741 gacatgctgc ctgctgatgc tctcgatgat ttcgatctgg atatgctccc ggccgacgcc
     1801 ctggacgact acgacctgga catcctcccg ggtaactaat aattaagtct cagccaccgt
     1861 taactgaaca tgtcaaaacc tgtggagact gttgagattt gatgttctga aaagataaag
     1921 cctataaata aaatgttgcc caaatttcct gcctgatgtt tttctttgtc tttgctacat
     1981 ggctttgctg ctcggatcgg ctcactctgt gtatgccacg ttcactttgt actctccttc
     2041 tcacggtagg tttattattt ttagatgtgc agttagtttc tgtgaaataa cacaccacac
     2101 actgatattg tctgtgcatt gacttggtga gtgcacattg tttttgatct tgacatattt
     2161 atatttgatt gatcaggtga actgtgtgaa tctaaagtgc tccatacaga tgttctgcat
     2221 tgaaaatatt ctcattttat tagtggaagt gagtgtatgc cacatccaat caatttcagc
     2281 aaacacccca gtatgattta atgcaaaaaa atgaaggtat caaacacgca ttactacttt
     2341 gcagttaaat atttaacatt tattccaaca cgaaaaaaag cagtaaataa cactttgaca
     2401 aacacgtcag gacatcttat ttttgtcacc ctcacaggca atttagtata atatattata
     2461 tatatatata tatcatataa taatattcag tataatatat atatatatat catattataa
     2521 tattcagtat aatataaaac acaaacacat atatgtataa tataatataa catttttatt
     2581 tattgagatg cctctatgga ccgtgttata agaagtaaag atcaggagaa gtaaacatga
     2641 agtgtaatta tgaatactga tgttaaatta agctatgatg agttttcact gttaatttac
     2701 catctcaatt aaatgttgat gcctccatga ccaagttaag cagatgagac tgagacaact
     2761 gtagaagaca agatgttcac tttgctgaat atagctggct tgacagttat ctatgactct
     2821 ataaatatat atatattttt ttttttataa aatgatttat ttataactat atatccattt
     2881 ctcagacagg tgcttcatat ccctcactcc cgtagctgtc catgctggat ctgtccccgt
     2941 tgtttttaaa aagctaaata agttattaac atgactgcat ccagcgagcc aaacctgtct
     3001 ggtgtacagc taccagagaa gcttgagatc ctagtcaaag ctttcaacca ctccaggcat
     3061 agctagagcc actgatgcat ctatagagct gaagtaaaga tagggttgat gcaaatcgat
     3121 ttagaatttt caatgtagaa aaaacaaaat aaaaagtcta ttcacttgat ggatagatta
     3181 gtcaaacaaa catagagtag gcacttccct tgactctgta aacttttgga tttaacagat
     3241 tgtgcactcc ttttagttaa acacattagt ccctgaaaac taagacccta accataatcc
     3301 cattgtagac ccacctactc gggtgaatat aaagctcgca cagatcatct cacagacatc
     3361 agaccttggc aaaggagcat cctgaacatc tctacacact gaggcaacga attcacctcg
     3421 agaccatggc tccaaagaag aagcgtaagg taaccatgag cgagctgatt aaggagaaca
     3481 tgcacatgaa gctgtacatg gagggcaccg tggacaacca tcacttcaag tgcacatccg
     3541 agggcgaagg caagccctac gagggcaccc agaccatgag aatcaaggtg gtcgagggcg
     3601 gccctctccc cttcgccttc gacatcctgg ctactagctt cctctacggc agcaagacct
     3661 tcatcaacca cacccagggc atccccgact tcttcaagca gtccttccct gagggcttca
     3721 catgggagag agtcaccaca tacgaagacg ggggcgtgct gaccgctacc caggacacca
     3781 gcctccagga cggctgcctc atctacaacg tcaagatcag aggggtgaac ttcacatcca
     3841 acggccctgt gatgcagaag aaaacactcg gctgggaggc cttcaccgag acgctgtacc
     3901 ccgctgacgg cggcctggaa ggcagaaacg acatggccct gaagctcgtg ggcgggagcc
     3961 atctgatcgc aaacatcaag accacatata gatccaagaa acccgctaag aacctcaaga
     4021 tgcctggcgt ctactatgtg gactacagac tggaaagaat caaggaggcc aacaacgaga
     4081 cctacgtcga gcagcacgag gtggcagtgg ccagatactg cgacctccct agcaaactgg
     4141 ggcacaagct caattaaagc tgagatctac ggactgttac cacttcacgc cgactcaaac
     4201 tgcgcagaga aacttcaaac gacaacattg gcatggcttt tgttattttt ggcgcttgac
     4261 tcaggatcta aaaactggaa cggtgaaggt gacggcaatg ttttttggca aataagcatc
     4321 cccgaagttc tacaatgcat ctgaggactc catgttttgt tttctttagt cattccaaat
     4381 gtttgttaaa tgcattgttc cgaaacttat ttgcctctat gaaggctgcc cagtaattgg
     4441 gagcatactt aacattgtag tattgtatgt aaattatgta acaaaacaat gtctgggttt
     4501 ttgtactttt cagccttaaa atcttgggtt tatttttttt tctttgttcc aaaaaaacga
     4561 agctttacca ttcaagatgt aaaggtttcc attccccctg ggcatattgt aaaagctgtg
     4621 tggaacgtgg cggtgccaga catttggtgg ggccaacctg tacactgact aattcaattc
     4681 caataaaagt gcacatgtaa gacatcctac ccctgtctca tttcctgtct tggctctgag
     4741 tggcttcaat aaaaatggct tttgtgaagt tgcaccatgt taattacatt tcagcacttt
     4801 cacaccaggt ttttgactaa tttgaattta tcgaagtttg accttttttt ttttttttct
     4861 tcaagctaat ttgaggttat caaaccattt cattggagga aacttaatgt ttaaaagcat
     4921 aaattcgaaa catgattcat ttatttaact gcttaatggt gaaacaaact tcaagtaata
     4981 caccaagtgt tccctcatat gaaggtagct aactgcaaac ggccttaact ttccatctta
     5041 agttgagcta cataaaggct ttaacaggaa ggaactgcca gtttaaaagt caaaccacca
     5101 tgactgatat aaattcttgt ccatataacc ttgcgtttca cacaatatag ctatctactg
     5161 ctaaataaat tagagctcat aacttcgtat aatgtatgct atacgaagtt atgttagata
     5221 acttcgtata aagtatccta tacgaagtta tgcgagctaa ctttaaataa ttggcattat
     5281 ttaaagttat gaatcgataa gtgaagagct atccggatat atatgtatgt tatcataact
     5341 gtactggcta tgtcaggtgg taatgactgt aacgttacgt tactcgttgt aggcacgaca
     5401 ttgaatatgc tcttctccgc tgtggcccga acgcctcccg ctagcgatgg gatctaatac
     5461 gactcactat agggagacca agggaaaata gaatgaagtg atctccaaaa aataagtact
     5521 ttttgactgt aaataaaatt gtaaggagta aaaagtactt ttttttctaa aaaaatgtaa
     5581 ttaagtaaaa gtaaaagtat tgatttttaa ttgtactcaa gtaaagtaaa aatccccaaa
     5641 aataatactt aagtacagta atcaagtaaa attactcaag tactttacac ctctgggatc
     5701 cagctgcatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
     5761 tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt
     5821 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct
     5881 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     5941 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     6001 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
     6061 gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     6121 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     6181 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     6241 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     6301 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     6361 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     6421 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     6481 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     6541 ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg taacactggc
     6601 aaaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6661 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6721 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6781 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6841 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6901 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6961 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     7021 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     7081 actgagatac ctacagcgtg agctatgaga aagcgccacg gttcccgaag ggagaaaggc
     7141 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     7201 gggaaacgcc tggtatcttt atagtcctgt
//
