LOCUS       Exported                7410 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 7410)
  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..7410
                     /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    3136..3150
                     /label=HR
     misc_feature    3136..3139
                     /label=last site
     misc_feature    3137..3217
                     /label=C-ATL-GLG
     misc_feature    3224..3313
                     /label=G1397-C
     CDS             3326..3574
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     polyA_signal    3611..3835
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    3825..3839
                     /label=HR
     promoter        3842..4352
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             4374..4973
                     /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    5120..5241
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(5660..6248)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6419..7279)
                     /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(7280..7384)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7400..7410
                     /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 ctagtcatga
     1441 cggtggcaaa caggctcttg aaaccgtcca acgccttcta ccagttctct gtcaagccca
     1501 cggactaacc ccagcgcagg ttgtagcgat tgctagtaat gggggtggca aacaggctct
     1561 tgagactgtt cagcgccttc taccagttct ctgtcaagcc cacggcctga cgcccgagca
     1621 agttgtagcg attgctagtc atgacggtgg caaacaggct cttgaaaccg tccaacgcct
     1681 tctaccagtt ctctgtcaag cccacggact aaccccagcg caagtggtag cgattgctag
     1741 taatattggt ggcaaacagg ctcttgagac ggttcagcgc ctccttccag ttctctgtca
     1801 agcccacgga ctcaccccag atcaagttgt agcgattgct agtaatgggg gtggcaaaca
     1861 ggctcttgag actgttcagc gccttctacc agttctctgt caagcccacg gcctgacgcc
     1921 cgagcaagtt gtagcgattg ctagtaatgg gggtggcaaa caggctcttg agactgttca
     1981 gcgccttcta ccagttctct gtcaagccca cggcctgacg cccgagcaag ttgtagcgat
     2041 tgctagtaat gggggtggca aacaggctct tgagactgtt cagcgccttc taccagttct
     2101 ctgtcaagcc cacggcctga cgcccgagca agttgtagca attgctagtc atgacggtgg
     2161 caaacaggct cttgaaaccg tccaacgcct tctaccagtt ctctgtcaag cccacggact
     2221 aaccccagcg caagttgtag cgattgctag taatattggt ggcaaacagg ctcttgagac
     2281 ggttcagcgc ctccttccag ttctctgtca agcccacgga ctcaccccag atcaagttgt
     2341 agcgattgct agtaatattg gtggcaaaca ggctcttgag acggttcagc gcctccttcc
     2401 agttctctgt caagcccacg gactcacccc agatcaagtt 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 tcatgacggt ggcaaacagg ctcttgaaac cgtccaacgc cttctaccag ttctctgtca
     2821 agcccacgga ctaaccccag cgcaagttgt agcgattgct agtaatattg gtggcaaaca
     2881 ggctcttgag acggttcagc gcctccttcc agttctctgt caagcccacg gactcacccc
     2941 agatcaagtt gtagcgattg cttccaatgg gggtggcaaa caggctcttg agactgttca
     3001 gcgccttcta ccagttctct gtcaagccca cggcctgacg cccgagcaag ttgtagcgat
     3061 tgcctctaat gggggcggca ggccggcgct ggagagcatt gttgcccagt tatctcgccc
     3121 tgatccggcg ttggccgcgt tgaccaacga ccacctcgtc gccttggcct gcctcggcgg
     3181 acgtcctgcg ctggatgcag tgaaaaaggg attgggaggc tccgcaatcc ctgtgaagag
     3241 aggcgccacc ggcgagacaa aggtgtttac cggcaactct aatagcccta agtccccaac
     3301 aaagggcggc tgctctggag gcagcaccaa cctgtctgac atcatcgaga aggagacagg
     3361 caagcagctg gtcatccagg agagcatcct gatgctgccc gaagaagtcg aagaagtgat
     3421 cggaaacaag cctgagagcg atatcctggt ccataccgcc tacgacgaga gtaccgacga
     3481 aaatgtgatg ctgctgacat ccgacgcccc agagtataag ccctgggctc tggtcatcca
     3541 ggattccaac ggagagaaca aaatcaaaat gctgctagag ggcccgttta aacccgctga
     3601 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     3661 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     3721 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3781 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggctcga
     3841 gggggttggg gttgcgcctt ttccaaggca gccctgggtt tgcgcaggga cgcggctgct
     3901 ctgggcgtgg ttccgggaaa cgcagcggcg ccgaccctgg gactcgcaca ttcttcacgt
     3961 ccgttcgcag cgtcacccgg atcttcgccg ctacccttgt gggccccccg gcgacgcttc
     4021 ctgctccgcc cctaagtcgg gaaggttcct tgcggttcgc ggcgtgccgg acgtgacaaa
     4081 cggaagccgc acgtctcact agtaccctcg cagacggaca gcgccaggga gcaatggcag
     4141 cgcgccgacc gcgatgggct gtggccaata gcggctgctc agcagggcgc gccgagagca
     4201 gcggccggga aggggcggtg cgggaggcgg ggtgtggggc ggtagtgtgg gccctgttcc
     4261 tgcccgcgcg gtgttccgca ttctgcaagc ctccggagcg cacgtcggca gtcggctccc
     4321 tcgttgaccg aatcaccgac ctctctcccc agggggatcc accggagctt accatgaccg
     4381 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     4441 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     4501 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     4561 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     4621 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     4681 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     4741 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     4801 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggaaacct
     4861 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     4921 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc
     4981 cccacgaccc gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg gtacctttaa
     5041 gaccaatgac ttacaaggca gctgtagatc ttagccactt tctagagtcg gggcggccgg
     5101 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     5161 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     5221 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     5281 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat
     5341 aaggatccgt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc
     5401 gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg
     5461 acaggtgccg gcagcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5521 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5581 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5641 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5701 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5761 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5821 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
     5881 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5941 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6001 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6061 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     6121 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6181 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6241 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6301 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6361 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6421 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6481 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6541 gtgctgcaat gataccgcgg gacccacgct caccggctcc agatttatca gcaataaacc
     6601 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6661 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6721 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6781 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6841 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6901 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6961 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7021 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7081 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7141 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7201 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7261 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7321 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7381 cgcgcacatt tccccgaaaa gtgccacctg
//
