LOCUS       Exported                4903 bp ds-DNA     circular SYN 22-FEB-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4903)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Feb 23, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4903
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     122..138
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    190..545
                     /label=LTF
     promoter        567..1066
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             1086..1685
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     CDS             1692..1748
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1749..2468
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    2496..2720
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    2742..2969
                     /label=RTF
     primer_bind     complement(3006..3022)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(3205..3793)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3964..4824)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLRNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERKRQIAEIGAS
                     LIKHW"
     promoter        complement(join(4825..4903,1..26))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
       61 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
      121 tgtaaaacga cggccagtcg aaccacgcaa tgcgtctcga tccgcagtgt cttgcgtctc
      181 taagctttac agtggtatga gaaagtttgg gcaccccctc tgaggttgtg taataatgga
      241 ctctgacttc acacaaaatt atcttggtgg catgtccttc atttgcccat aagagccaag
      301 tgttagcttg atttccaaac acagatttta agtattcata gtatgcaatt gtatgaaatt
      361 aaatcaaatg taaaaaatag gctgtgcaaa aatgtgggaa cccttgtcat tttgttgctt
      421 tgatggcctg taactaatca tcacaagata actggacaca caaagttggt ttggtgagct
      481 cgttaagcct tgaacttaat tgacaggtgc ataaaataat gacaaaaggt atttaaggtg
      541 gccaaaccgg tgctagcaat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg
      601 agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg
      661 cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc
      721 gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg
      781 caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc
      841 tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc
      901 tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag
      961 gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg
     1021 cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcgacct gcagccaacg
     1081 ccaccatggg gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc
     1141 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg
     1201 acccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg
     1261 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca
     1321 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt
     1381 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc
     1441 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg
     1501 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg
     1561 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg
     1621 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg
     1681 gtgccatcga tgccactaac ttctccctgt tgaaacaagc aggggatgtc gaagagaatc
     1741 ccgggccaat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     1801 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     1861 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     1921 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     1981 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     2041 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     2101 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     2161 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     2221 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     2281 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     2341 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     2401 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     2461 acaagtaatt aattaagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     2521 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2581 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     2641 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     2701 ggatgcggtg ggctctatgg cttctgaggc gcgccctcga gtatgatttt tctattaagg
     2761 tgcccaaatt tatgcacagg cctttttttg ttttggggca tattgcacat tttctgttaa
     2821 cccaataaac ctcatttcac tgctgaaata tgattgtgtt catcagttat ttgatatatc
     2881 aaaattaagt tgtgaattaa aacacccaat gattggtgaa taaaaataat gccaattgtc
     2941 aagggtgccc aaactttctc ataccactgt agggcccaga gacggagtca ctgccaaccg
     3001 agacggtcat agctgtttcc tgtgtgccgc ttcctcgctc actgactcgc tgcgctcggt
     3061 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tacccacaga
     3121 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3181 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3241 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3301 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3361 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3421 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3481 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3541 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3601 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3661 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3721 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3781 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3841 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3901 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3961 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctct ttcgttcatc
     4021 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4081 ccccagtgct gcaataatac cgcgggaccc acgctcaccg gctccagatt tatcagcaat
     4141 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4201 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4261 caacgttgtt gccatcgcta caggcatcgt ggtatcacgc tcgtcgtttg gtatggcttc
     4321 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgcgcaaaaa
     4381 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg ccgtgttatc
     4441 actcatggtt atggcagcac tacataattc tcttactgtc atgccatccg taagatgctt
     4501 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4561 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4621 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4681 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4741 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4801 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4861 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tag
//
