LOCUS       Exported                3491 bp DNA     circular SYN 20-AUG-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3491)
  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..3491
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    21..64
                     /label=HBB 5'UTR
     primer_bind     80..127
                     /label=Puro_GA-OL_Fwd
     misc_feature    101..106
                     /label=Kozak
     primer_bind     105..125
                     /label=PuroR_GA-OL_Fwd
     CDS             107..706
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MATEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGSRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     606..626
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(688..727)
                     /label=Puro_F2A-GA-OL_Fwd
     primer_bind     complement(699..715)
                     /label=Puro-GSG_Rv
     primer_bind     706..730
                     /label=F2A_BleoR_GA-OL_Fwd
     CDS             716..781
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     primer_bind     734..798
                     /label=F2A_Clover3_GA-OL-GA2_Fwd
     primer_bind     complement(766..784)
                     /label=T2Aatg_Up_Rv
     primer_bind     780..799
                     /label=Kozak-mClover3_SacI_Fwd
     primer_bind     780..794
                     /label=Kozak-mClover3_BamHI_Fwd
     CDS             782..1501
                     /codon_start=1
                     /label=mClover3
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNCIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(1478..1538)
                     /label=Clover3_GA-OL-GA3_Rv
     primer_bind     complement(1479..1501)
                     /label=mClover_XbaI_Rv
     primer_bind     1538..1564
                     /label=HBB_3'UTR_Fwd
     3'UTR           1539..1672
                     /label=HBB 3'UTR
     primer_bind     complement(1539..1557)
                     /label=HBB_3UTR_seq_Rv
     primer_bind     1673..1701
                     /label=pUC19_PciI-to-NcoI_Fwd
     rep_origin      complement(1736..2324)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2495..3355)
                     /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(3356..3460)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3468..3491)
                     /label=pUC_AatII_up
ORIGIN
        1 taatacgact cactataggg acatttgctt ctgacacaac tgtgttcact agcaacctca
       61 aacagaattc gagctcggta cccggggatc ctctagagtc gccaccatgg ccaccgagta
      121 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc
      181 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga
      241 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
      301 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
      361 gggggcggtg ttcgccgaga tcggctcgcg catggccgag ttgagcggtt cccggctggc
      421 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
      481 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
      541 gctccccgga gtggaggcgg ccgagcgcgc tggggtgccc gccttcctgg agacctccgc
      601 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
      661 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgccggca gtggagtgaa
      721 acagactttg aattttgacc ttctcaagtt ggcgggagac gtcgagtcca accctgggcc
      781 catggtgtct aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
      841 cggcgacgta aacggccaca agttcagcgt ccgcggcgag ggcgagggcg atgccaccaa
      901 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
      961 cctcgtgacc accttcggct acggcgtggc ctgcttcagc cgctaccccg accacatgaa
     1021 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctc
     1081 tttcaaggac gacggtacct acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     1141 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1201 caagctggag tacaacttca acagccacta cgtctatatc acggccgaca agcagaagaa
     1261 ctgcatcaag gctaacttca agatccgcca caacgttgag gacggcagcg tgcagctcgc
     1321 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1381 ctacctgagc catcagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt
     1441 cctgctggag ttcgtgaccg ccgccgggat tacacatggc atggacgagc tgtacaagta
     1501 agttcgcgtc tggaacagac ctgcaggcat gcaagcttgc tcgctttctt gctgtccaat
     1561 ttctattaaa ggttcctttg ttccctaagt ccaactacta aactggggga tattatgaag
     1621 ggccttgagc atctggattc tgcctaataa aaaacattta ttttcattgc aaaaccatgg
     1681 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1741 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1801 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     1861 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     1921 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     1981 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2041 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2101 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2161 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2221 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2281 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2341 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2401 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2461 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     2521 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     2581 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     2641 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     2701 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     2761 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     2821 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     2881 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     2941 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3001 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3061 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3121 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     3181 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     3241 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     3301 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     3361 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     3421 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     3481 cacctgacgt c
//
