LOCUS       Exported                7296 bp ds-DNA     circular SYN 21-FEB-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7296)
  AUTHORS   Triple Threat
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Feb 22, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7296
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..4
                     /label=HR
     enhancer        8..387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        388..591
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        636..654
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    654..668
                     /label=HR
     CDS             674..853
                     /codon_start=1
                     /label=MTS
                     /translation="MLGFVGRVAAAPASGALRRLTPSASLPPAQLLLRAAPTAVHPVRD
                     YAAQTSESGGGGSPG"
     CDS             863..886
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    882..896
                     /label=HR
     CDS             893..1300
                     /codon_start=1
                     /label=N136aa
                     /translation="VDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALS
                     QHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLD
                     TGQLLKIAKRGGVTAVEAVHAWRNALTGAPLN"
     misc_feature    1220..1222
                     /label=N1
     misc_feature    1229..1231
                     /label=N2
     misc_feature    1232..1234
                     /label=N3
     misc_feature    1305..1308
                     /label=1st site
     misc_feature    2830..2844
                     /label=HR
     misc_feature    2830..2833
                     /label=last site
     misc_feature    2831..2911
                     /label=C-ATL-GLG
     misc_feature    2918..3199
                     /label=G1333C
     CDS             3212..3460
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     polyA_signal    3497..3721
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    3711..3725
                     /label=HR
     promoter        3728..4238
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             4260..4859
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    5006..5127
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(5546..6134)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6305..7165)
                     /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(7166..7270)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7286..7296
                     /label=HR
ORIGIN
        1 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
       61 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      181 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      241 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      301 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      361 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      421 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      481 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      541 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
      601 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagaca
      661 agctggctag accatgttgg ggtttgtggg tcgggtggcc gctgctccgg cctccggggc
      721 cttgcggaga ctcacccctt cagcgtcgct gcccccagct cagctcttac tgcgggccgc
      781 tccgacggcg gtccatcctg tcagggacta tgcggcgcaa acatctgaga gcggcggagg
      841 cgggagcccc ggcgcggccg ctgactacaa ggatgacgac gataaaggta gcgtggatct
      901 acgcacgctc ggctacagcc agcagcaaca ggagaagatc aaaccgaagg ttcgttcgac
      961 agtggcgcag caccacgagg cactggtcgg ccatgggttt acacacgcgc acatcgttgc
     1021 gctcagccaa cacccggcag cgttagggac cgtcgctgtc aagtatcagg acatgatcgc
     1081 agcgttgcca gaggcgacac acgaagcgat cgttggcgtc ggcaaacagt ggtccggcgc
     1141 acgcgctctg gaggccttgc tcacggtggc gggagagttg agaggtccac cgttacagtt
     1201 ggacacaggc caacttctca agattgcaaa acgtggcggc gtgaccgcag tggaggcagt
     1261 gcatgcatgg cgcaatgcac tgacgggtgc ccccctgaac ctgaccccgg agcaggtggt
     1321 ggccatcgct agtcatgacg gtggcaaaca ggctcttgaa accgtccaac gccttctacc
     1381 agttctctgt caagcccacg gactaacccc agcgcaagtt gtagcgattg ctagtaatgg
     1441 gggtggcaaa caggctcttg agactgttca gcgccttcta ccagttctct gtcaagccca
     1501 cggcctgacg cccgagcagg ttgtagcgat tgctagtaat attggtggca aacaggctct
     1561 tgagacggtt cagcgcctcc ttccagttct ctgtcaagcc cacggactca ccccagatca
     1621 agttgtagcg attgctagtc atgacggtgg caaacaggct cttgaaaccg tccaacgcct
     1681 tctaccagtt ctctgtcaag cccacggact aaccccagcg caagtggtag cgattgctag
     1741 tcatgacggt ggcaaacagg ctcttgaaac cgtccaacgc cttctaccag ttctctgtca
     1801 agcccacgga ctaaccccag cgcaagttgt agcgattgct agtaatgggg gtggcaaaca
     1861 ggctcttgag actgttcagc gccttctacc agttctctgt caagcccacg gcctgacgcc
     1921 cgagcaagtt gtagcgattg ctagtaatat tggtggcaaa caggctcttg agacggttca
     1981 gcgcctcctt ccagttctct gtcaagccca cggactcacc ccagatcaag ttgtagcgat
     2041 tgctagtaat gggggtggca aacaggctct tgagactgtt cagcgccttc taccagttct
     2101 ctgtcaagcc cacggcctga cgcccgagca agttgtagca attgctagta atgggggtgg
     2161 caaacaggct cttgagactg ttcagcgcct tctaccagtt ctctgtcaag cccacggcct
     2221 gacgcccgag caagttgtag cgattgctag taatattggt ggcaaacagg ctcttgagac
     2281 ggttcagcgc ctccttccag ttctctgtca agcccacgga ctcaccccag atcaagttgt
     2341 agcgattgct agtaatgggg gtggcaaaca ggctcttgag actgttcagc gccttctacc
     2401 agttctctgt caagcccacg gcctgacgcc cgagcaagtt gtagcgattg ctagtaatgg
     2461 gggtggcaaa caggctcttg agactgttca gcgccttcta ccagttctct gtcaagccca
     2521 cggcctgacg cccgagcaag ttgtagcgat agccagtaat gggggtggca aacaggctct
     2581 tgagactgtt cagcgccttc taccagttct ctgtcaagcc cacggcctga cgcccgagca
     2641 agttgtagcg attgctagta acaatggtgg caaacaggct cttgaaaccg tacagcgcct
     2701 actgccagtt ctctgtcaag cccacggtct gactccggag caagttgtag cgattgcaag
     2761 taacaatggc ggcaggccgg cgctggagag cattgttgcc cagttatctc gccctgatcc
     2821 ggcgttggcc gcgttgacca acgaccacct cgtcgccttg gcctgcctcg gcggacgtcc
     2881 tgcgctggat gcagtgaaaa agggattggg aggctcccca accccctacc ctaactatgc
     2941 caatgcagga cacgtggagg gacagagcgc cctgttcatg cgggataacg gcatctccga
     3001 gggcctggtg ttccacaaca atcccgaggg cacctgcggc ttttgcgtga acatgaccga
     3061 gacactgctg cccgagaatg ccaagatgac agtggtgcca cctgagggag caatccctgt
     3121 gaagagaggc gccaccggcg agacaaaggt gtttaccggc aactctaata gccctaagtc
     3181 cccaacaaag ggcggctgct ctggaggcag caccaacctg tctgacatca tcgagaagga
     3241 gacaggcaag cagctggtca tccaggagag catcctgatg ctgcccgaag aagtcgaaga
     3301 agtgatcgga aacaagcctg agagcgatat cctggtccat accgcctacg acgagagtac
     3361 cgacgaaaat gtgatgctgc tgacatccga cgccccagag tataagccct gggctctggt
     3421 catccaggat tccaacggag agaacaaaat caaaatgctg ctagagggcc cgtttaaacc
     3481 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3541 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3601 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3661 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3721 gctcgagggg gttggggttg cgccttttcc aaggcagccc tgggtttgcg cagggacgcg
     3781 gctgctctgg gcgtggttcc gggaaacgca gcggcgccga ccctgggact cgcacattct
     3841 tcacgtccgt tcgcagcgtc acccggatct tcgccgctac ccttgtgggc cccccggcga
     3901 cgcttcctgc tccgccccta agtcgggaag gttccttgcg gttcgcggcg tgccggacgt
     3961 gacaaacgga agccgcacgt ctcactagta ccctcgcaga cggacagcgc cagggagcaa
     4021 tggcagcgcg ccgaccgcga tgggctgtgg ccaatagcgg ctgctcagca gggcgcgccg
     4081 agagcagcgg ccgggaaggg gcggtgcggg aggcggggtg tggggcggta gtgtgggccc
     4141 tgttcctgcc cgcgcggtgt tccgcattct gcaagcctcc ggagcgcacg tcggcagtcg
     4201 gctccctcgt tgaccgaatc accgacctct ctccccaggg ggatccaccg gagcttacca
     4261 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
     4321 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
     4381 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     4441 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     4501 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     4561 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     4621 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     4681 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     4741 aaacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     4801 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac
     4861 gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc acgaccccat gcatcggtac
     4921 ctttaagacc aatgacttac aaggcagctg tagatcttag ccactttcta gagtcggggc
     4981 ggccggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
     5041 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     5101 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     5161 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa
     5221 atcgataagg atccgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg
     5281 gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact
     5341 cgtaggacag gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5401 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5461 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5521 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5581 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5641 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5701 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     5761 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5821 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5881 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5941 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     6001 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6061 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6121 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6181 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6241 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6301 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6361 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6421 gccccagtgc tgcaatgata ccgcgggacc cacgctcacc ggctccagat ttatcagcaa
     6481 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6541 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6601 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6661 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6721 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6781 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6841 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6901 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6961 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7021 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7081 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7141 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7201 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7261 gggttccgcg cacatttccc cgaaaagtgc cacctg
//
