LOCUS       Exported                6214 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 6214)
  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..6214
                     /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    195..1387
                     /label=LTF
     promoter        1409..1908
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             1928..2527
                     /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             2534..2590
                     /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             2591..3310
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    3338..3562
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    3584..4275
                     /label=RTF
     primer_bind     complement(4317..4333)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(4516..5104)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5275..6135)
                     /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(6136..6214,1..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 tcatatggtc ctggcaaaga cactagaata acaagatgcg taacgccata cgattttttg
      241 gcacacgatt ttttcgccgt ggctctagag gtggctccag gctctctcga atttttgttc
      301 gagagcgaga gagcgaagag cgctacagcg aacagctctt ttctacgcat acagtgatag
      361 cagacaactg tatgtgtgca cacgtatgct catgcattgt aaatttgaca aaatatgccc
      421 ttcaccttag aagttcttag actttaaatc tatattattt ttgatcaatt ggcaccatgc
      481 gaaaaattct tgttttgcat tgccttaacg ttattattat ttgaaaatag attagaaata
      541 gccaaatcta tgtacatatt atcacaaaat aaatttcaaa aatgacttta tattagaata
      601 tttgtcatta gagtattcag cttgcggcgt gtgaaaaatt aataaggcaa tgattgttga
      661 gtgcttgtgt ccgcacttcg tgcctcaaga tatgaccaaa gcaaagacac tagaataatt
      721 ctagtgtctt tgatatgact tttgcaataa acagttttca tatttttatt tattttataa
      781 atttttattt tctacttcgt attattttta tgaaatattt atttctcaat gtaatgtctt
      841 ctttttgtaa aatttatgtt ttaaattaca gtagttataa taatttatat tttacttcct
      901 ttaattattt agtatattta ttaagtcatt tgacttaata tgatttatgt atgtaaatga
      961 tccattttta ttatttaaaa tgcacattag attcagttgt ttttcagttt gttttttttt
     1021 tttgttatat tccaattcga tatttttaaa aagcgcgcca tttcgggtgg gaagaaaagc
     1081 gcgccaattt ttttgttttt ccttttaatt tctcttagcc cgtttgctta aagttggcta
     1141 tacgcagagc tcctcgattt tttgatattt tcgctatata atcgaattta tattcaacta
     1201 aagtcgtttt gttgcaaaag taaagtacgg aaatataagc gcaaatagta attgcgataa
     1261 atagtgaata gtgaatagtg tttacgtatt ttcgtaattt gacgcgctaa tacaacgtgc
     1321 tgctgctttt cgacaaaggt ttgagttgaa atttataaat tgtaagtgaa aaaaaaaaaa
     1381 cattttaacc ggtgctagca attctaccgg gtaggggagg cgcttttccc aaggcagtct
     1441 ggagcatgcg ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct
     1501 cgcacacatt ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc
     1561 gcgccacctt ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg
     1621 tgcaggacgt gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc
     1681 gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc
     1741 gctttctggg ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggctc
     1801 aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag
     1861 cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcagccaa
     1921 cgccaccatg gggaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     1981 ccgggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     2041 cgacccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     2101 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     2161 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     2221 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     2281 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     2341 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     2401 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     2461 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     2521 cggtgccatc gatgccacta acttctccct gttgaaacaa gcaggggatg tcgaagagaa
     2581 tcccgggcca atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     2641 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     2701 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     2761 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     2821 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     2881 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     2941 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     3001 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     3061 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     3121 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     3181 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     3241 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     3301 gtacaagtaa ttaattaagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
     3361 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3421 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3481 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     3541 ggggatgcgg tgggctctat ggcttctgag gcgcgccctc gagaaaaaat gagcgatatt
     3601 taaataactg acgactgaaa aatcgagcaa tatataataa aaaacaatag ttagaaaaat
     3661 aacttaattg gggaacatgg aaatgtttta atgttaaaca ttaaaatatt tcaagtcgac
     3721 tcgaaggtca tatacgtaaa tataggaccc cattacaatt gtaatggcct ccccgtggtg
     3781 ttccctgggt accgattatt acacatatta cacataatta attatcaaca taaatataaa
     3841 tatgtaaacg gtagctaatt cgagcggcga ttttaacaaa cgaatttaaa aagctttaaa
     3901 attaaaaaat tttaaaatta aaattaaaat tttaaaatta taataatcag ggcgcgaatt
     3961 tttaaaaatt attttatttc atcatattgc tacgaaattg gcaaaaaact accctaatat
     4021 gtacaatgta aattcatttc ttcgatcaga attgatttcg gcccgaaaat cgtcttctag
     4081 cacaacacgc acacatatac gcgttctcgt ctcttgtttt tactcacaca agcaagcaaa
     4141 ttctattttt agatttctta cgttctcagc gtgagcgagc ggaaagagag caattttggc
     4201 cgtcaccaaa aaagtggctg catagtgcca aaccaatgta tggccgttac gcatcttgtt
     4261 attctagtgt ctttggtcct gggggcccag agacggagtc actgccaacc gagacggtca
     4321 tagctgtttc ctgtgtgccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4381 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttacccacag aatcagggga
     4441 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4501 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4561 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4621 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4681 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4741 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4801 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4861 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4921 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4981 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5041 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5101 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5161 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5221 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5281 atgcttaatc agtgaggcac ctatctcagc gatctgtctc tttcgttcat ccatagttgc
     5341 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5401 tgcaataata ccgcgggacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5461 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5521 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5581 tgccatcgct acaggcatcg tggtatcacg ctcgtcgttt ggtatggctt cattcagctc
     5641 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgcgcaaaa aagcggttag
     5701 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gccgtgttat cactcatggt
     5761 tatggcagca ctacataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5821 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5881 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5941 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6001 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6061 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6121 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6181 tctcatgagc ggatacatat ttgaatgtat ttag
//
