LOCUS       Exported                7302 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 7302)
  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..7302
                     /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    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    2924..3205
                     /label=G1333C
     CDS             3218..3466
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     polyA_signal    3503..3727
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    3717..3731
                     /label=HR
     promoter        3734..4244
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             4266..4865
                     /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    5012..5133
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(5552..6140)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6311..7171)
                     /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(7172..7276)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7292..7302
                     /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 agtaatattg gtggcaaaca ggctcttgag acggttcagc gcctccttcc
     1381 agttctctgt caagcccacg gactcacccc agatcaagtt gtagcgattg ctagtcatga
     1441 cggtggcaaa caggctcttg aaaccgtcca acgccttcta ccagttctct gtcaagccca
     1501 cggactaacc ccagcgcagg ttgtagcgat tgctagtaat gggggtggca aacaggctct
     1561 tgagactgtt cagcgccttc taccagttct ctgtcaagcc cacggcctga cgcccgagca
     1621 agttgtagcg attgctagta acaatggtgg caaacaggct cttgaaaccg tacagcgcct
     1681 actgccagtt ctctgtcaag cccacggtct gactccggag caagtggtag cgattgctag
     1741 taatattggt ggcaaacagg ctcttgagac ggttcagcgc ctccttccag ttctctgtca
     1801 agcccacgga ctcaccccag atcaagttgt agcgattgct agtaacaatg gtggcaaaca
     1861 ggctcttgaa accgtacagc gcctactgcc agttctctgt caagcccacg gtctgactcc
     1921 ggagcaagtt gtagcgattg ctagtaacaa tggtggcaaa caggctcttg aaaccgtaca
     1981 gcgcctactg ccagttctct gtcaagccca cggtctgact ccggagcaag ttgtagcgat
     2041 tgctagtaat attggtggca aacaggctct tgagacggtt cagcgcctcc ttccagttct
     2101 ctgtcaagcc cacggactca ccccagatca agttgtagca attgctagta acaatggtgg
     2161 caaacaggct cttgaaaccg tacagcgcct actgccagtt ctctgtcaag cccacggtct
     2221 gactccggag caagttgtag cgattgctag taatgggggt ggcaaacagg ctcttgagac
     2281 tgttcagcgc cttctaccag ttctctgtca agcccacggc ctgacgcccg agcaagttgt
     2341 agcgattgct agtaatattg gtggcaaaca ggctcttgag acggttcagc gcctccttcc
     2401 agttctctgt caagcccacg gactcacccc agatcaagtt gtagcgattg ctagtcatga
     2461 cggtggcaaa caggctcttg aaaccgtcca acgccttcta ccagttctct gtcaagccca
     2521 cggactaacc ccagcgcaag ttgtagcgat agccagtcat gacggtggca aacaggctct
     2581 tgaaaccgtc caacgccttc taccagttct ctgtcaagcc cacggactaa ccccagcgca
     2641 agttgtagcg attgctagtc atgacggtgg caaacaggct cttgaaaccg tccaacgcct
     2701 tctaccagtt ctctgtcaag cccacggact aaccccagcg caagttgtag cgattgcaag
     2761 taacaatggc ggcaggccgg cgctggagag cattgttgcc cagttatctc gccctgatcc
     2821 ggcgttggcc gcgttgacca acgaccacct cgtcgccttg gcctgcctcg gcggacgtcc
     2881 tgcgctggat gcagtgaaaa agggattggg aggctccgct agcccaaccc cctaccctaa
     2941 ctatgccaat gcaggacacg tggagggaca gagcgccctg ttcatgcggg ataacggcat
     3001 ctccgagggc ctggtgttcc acaacaatcc cgagggcacc tgcggctttt gcgtgaacat
     3061 gaccgagaca ctgctgcccg agaatgccaa gatgacagtg gtgccacctg agggagcaat
     3121 ccctgtgaag agaggcgcca ccggcgagac aaaggtgttt accggcaact ctaatagccc
     3181 taagtcccca acaaagggcg gctgctctgg aggcagcacc aacctgtctg acatcatcga
     3241 gaaggagaca ggcaagcagc tggtcatcca ggagagcatc ctgatgctgc ccgaagaagt
     3301 cgaagaagtg atcggaaaca agcctgagag cgatatcctg gtccataccg cctacgacga
     3361 gagtaccgac gaaaatgtga tgctgctgac atccgacgcc ccagagtata agccctgggc
     3421 tctggtcatc caggattcca acggagagaa caaaatcaaa atgctgctag agggcccgtt
     3481 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     3541 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     3601 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     3661 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     3721 tctatggctc gagggggttg gggttgcgcc ttttccaagg cagccctggg tttgcgcagg
     3781 gacgcggctg ctctgggcgt ggttccggga aacgcagcgg cgccgaccct gggactcgca
     3841 cattcttcac gtccgttcgc agcgtcaccc ggatcttcgc cgctaccctt gtgggccccc
     3901 cggcgacgct tcctgctccg cccctaagtc gggaaggttc cttgcggttc gcggcgtgcc
     3961 ggacgtgaca aacggaagcc gcacgtctca ctagtaccct cgcagacgga cagcgccagg
     4021 gagcaatggc agcgcgccga ccgcgatggg ctgtggccaa tagcggctgc tcagcagggc
     4081 gcgccgagag cagcggccgg gaaggggcgg tgcgggaggc ggggtgtggg gcggtagtgt
     4141 gggccctgtt cctgcccgcg cggtgttccg cattctgcaa gcctccggag cgcacgtcgg
     4201 cagtcggctc cctcgttgac cgaatcaccg acctctctcc ccagggggat ccaccggagc
     4261 ttaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
     4321 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
     4381 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     4441 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     4501 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     4561 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     4621 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     4681 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     4741 tcctggaaac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     4801 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     4861 cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga ccccatgcat
     4921 cggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttctagagt
     4981 cggggcggcc ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     5041 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     5101 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     5161 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     5221 ggtaaaatcg ataaggatcc gtcgaccgat gcccttgaga gccttcaacc cagtcagctc
     5281 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat
     5341 gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc
     5401 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5461 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5521 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5581 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5641 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5701 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta
     5761 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5821 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5881 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5941 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     6001 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6061 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6121 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6181 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6241 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6301 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6361 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6421 catctggccc cagtgctgca atgataccgc gggacccacg ctcaccggct ccagatttat
     6481 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6541 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6601 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6661 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6721 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6781 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6841 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6901 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6961 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7021 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7081 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7141 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7201 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7261 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tg
//
