LOCUS       Exported                2779 bp DNA     circular SYN 04-JUN-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2779)
  AUTHORS   Carles Calatayud
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jun 4, 2021 from SnapGene Viewer 5.3.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..2779
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     18..42
                     /label=pIVTR(AG)(HBB)_Fwd
     misc_feature    21..64
                     /label=HBB 5'UTR
     primer_bind     64..88
                     /label=mScarlet_EcoRI_Fwd
     misc_feature    65
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     70..94
                     /label=T2A-mScarlet-I_GA-OL_Fwd
     primer_bind     70..92
                     /label=mScarlet_KpnI_Fwd
     regulatory      71..80
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             77..772
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(210..228)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     478..497
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(751..775)
                     /label=mScarlet-I_Rv
     primer_bind     complement(752..781)
                     /label=EBFP_SacI_Rv
     misc_feature    776
                     /label=3'UTR
     misc_feature    776
                     /label=pRR_DARPP32
     misc_feature    781..826
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     3'UTR           827..960
                     /label=HBB 3'UTR
     primer_bind     complement(827..845)
                     /label=HBB_3UTR_seq_Rv
     primer_bind     complement(924..962)
                     /label=HBB_3UTR_150T_Rv
     primer_bind     961..989
                     /label=pUC19_PciI-to-NcoI_Fwd
     rep_origin      complement(1024..1612)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1783..2643)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2644..2748)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2750..2769
                     /label=Pre_T7_Fwd
     primer_bind     complement(2756..2779)
                     /label=pUC_AatII_up
     primer_bind     complement(join(2769..2779,1..17))
                     /label=pIVTR(AG)(HBB)_Rv
     primer_bind     join(2775..2779,1..41)
                     /label=T7(AG)_IVTR_Fwd
ORIGIN
        1 taatacgact cactataagg acatttgctt ctgacacaac tgtgttcact agcaacctca
       61 aacagaattc gccaccatgg tgagcaaggg cgaggcagtg atcaaggagt tcatgcggtt
      121 caaggtgcac atggagggct ccatgaacgg ccacgagttc gagatcgagg gcgagggcga
      181 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct
      241 gcccttctcc tgggacatcc tgtcccctca gttcatgtac ggctccaggg ccttcaccaa
      301 gcaccccgcc gacatccccg actactataa gcagtccttc cccgagggct tcaagtggga
      361 gcgcgtgatg aacttcgagg acggcggcgc cgtgaccgtg acccaggaca cctccctgga
      421 ggacggcacc ctgatctaca aggtgaagct ccgcggcacc aacttccctc ctgacggccc
      481 cgtaatgcag aagaagacaa tgggctggga agcgtccacc gagcggttgt accccgagga
      541 cggcgtgctg aagggcgaca ttaagatggc cctgcgcctg aaggacggcg gccgctacct
      601 ggcggacttc aagaccacct acaaggccaa gaagcccgtg cagatgcccg gcgcctacaa
      661 cgtcgaccgc aagttggaca tcacctccca caacgaggac tacaccgtgg tggaacagta
      721 cgaacgctcc gagggccgcc actccaccgg cggcatggac gagctgtaca agtaagagct
      781 cggtacccgg ggatcctcta gagtcgacct gcaggcatgc aagcttgctc gctttcttgc
      841 tgtccaattt ctattaaagg ttcctttgtt ccctaagtcc aactactaaa ctgggggata
      901 ttatgaaggg ccttgagcat ctggattctg cctaataaaa aacatttatt ttcattgcaa
      961 aaccatggga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1021 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1081 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1141 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1201 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1261 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1321 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1381 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1441 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     1501 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     1561 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     1621 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     1681 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     1741 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     1801 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     1861 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     1921 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1981 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2041 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2101 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     2161 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     2221 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     2281 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     2341 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     2401 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     2461 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     2521 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     2581 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     2641 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     2701 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     2761 aaaagtgcca cctgacgtc
//
