LOCUS       Exported                7752 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 7752)
  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..7752
                     /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    3238..3252
                     /label=HR
     misc_feature    3238..3241
                     /label=last site
     misc_feature    3239..3319
                     /label=C-ATL-GLG
     misc_feature    3332..3655
                     /label=G1397-N
     CDS             3668..3916
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     polyA_signal    3953..4177
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    4167..4181
                     /label=HR
     promoter        4184..4694
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             4716..5315
                     /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    5462..5583
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(6002..6590)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6761..7621)
                     /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(7622..7726)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7742..7752
                     /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 agtaacaatg gtggcaaaca ggctcttgaa accgtacagc gcctactgcc
     1381 agttctctgt caagcccacg gtctgactcc ggagcaagtt 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 atgggggtgg caaacaggct cttgagactg ttcagcgcct
     1681 tctaccagtt ctctgtcaag cccacggcct gacgcccgag caagtggtag cgattgctag
     1741 taatgggggt ggcaaacagg ctcttgagac tgttcagcgc cttctaccag ttctctgtca
     1801 agcccacggc ctgacgcccg agcaagttgt 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 tgctagtcat gacggtggca aacaggctct tgaaaccgtc caacgccttc taccagttct
     2101 ctgtcaagcc cacggactaa ccccagcgca agttgtagca attgctagta acaatggtgg
     2161 caaacaggct cttgaaaccg tacagcgcct actgccagtt ctctgtcaag cccacggtct
     2221 gactccggag caagttgtag cgattgctag taacaatggt ggcaaacagg ctcttgaaac
     2281 cgtacagcgc ctactgccag ttctctgtca agcccacggt ctgactccgg agcaagttgt
     2341 agcgattgct agtaatattg gtggcaaaca ggctcttgag acggttcagc gcctccttcc
     2401 agttctctgt caagcccacg gactcacccc agatcaagtt gtagcgattg ctagtaacaa
     2461 tggtggcaaa caggctcttg aaaccgtaca gcgcctactg ccagttctct gtcaagccca
     2521 cggtctgact ccggagcaag ttgtagcgat agccagtaat gggggtggca aacaggctct
     2581 tgagactgtt cagcgccttc taccagttct ctgtcaagcc cacggcctga cgcccgagca
     2641 agttgtagcg attgctagta atattggtgg caaacaggct cttgagacgg ttcagcgcct
     2701 ccttccagtt ctctgtcaag cccacggact caccccagat caagttgtag cgattgcaag
     2761 taatgggggt ggcaaacagg ctcttgagac tgttcagcgc cttctaccag ttctctgtca
     2821 agcccacggc ctgacgcccg agcaagttgt agcgattgct agtaatattg gtggcaaaca
     2881 ggctcttgag acggttcagc gcctccttcc agttctctgt caagcccacg gactcacccc
     2941 agatcaagtt gtagcgattg cttccaatat tggtggcaaa caggctcttg agacggttca
     3001 gcgcctcctt ccagttctct gtcaagccca cggactcacc ccagatcaag ttgtagcgat
     3061 tgctagtaac aatggtggca aacaggctct tgaaaccgta cagcgcctac tgccagttct
     3121 ctgtcaagcc cacggtctga ctccggagca agttgtagcg attgcctcta atgggggcgg
     3181 caggccggcg ctggagagca ttgttgccca gttatctcgc cctgatccgg cgttggccgc
     3241 gttgaccaac gaccacctcg tcgccttggc ctgcctcggc ggacgtcctg cgctggatgc
     3301 agtgaaaaag ggattgggag gctccgctag cggctcctac gccctgggcc catatcagat
     3361 ctctgcccca cagctgccag catacaacgg acagaccgtg ggcacattct actatgtgaa
     3421 tgacgccggc ggcctggagt ctaaggtgtt tagctccggc ggcccaaccc cctaccctaa
     3481 ctatgccaat gcaggacacg tggagggaca gagcgccctg ttcatgcggg ataacggcat
     3541 ctccgagggc ctggtgttcc acaacaatcc cgagggcacc tgcggctttt gcgtgaacat
     3601 gaccgagaca ctgctgcccg agaatgccaa gatgacagtg gtgccacctg agggatctgg
     3661 aggcagcacc aacctgtctg acatcatcga gaaggagaca ggcaagcagc tggtcatcca
     3721 ggagagcatc ctgatgctgc ccgaagaagt cgaagaagtg atcggaaaca agcctgagag
     3781 cgatatcctg gtccataccg cctacgacga gagtaccgac gaaaatgtga tgctgctgac
     3841 atccgacgcc ccagagtata agccctgggc tctggtcatc caggattcca acggagagaa
     3901 caaaatcaaa atgctgctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     3961 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     4021 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     4081 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     4141 aatagcaggc atgctgggga tgcggtgggc tctatggctc gagggggttg gggttgcgcc
     4201 ttttccaagg cagccctggg tttgcgcagg gacgcggctg ctctgggcgt ggttccggga
     4261 aacgcagcgg cgccgaccct gggactcgca cattcttcac gtccgttcgc agcgtcaccc
     4321 ggatcttcgc cgctaccctt gtgggccccc cggcgacgct tcctgctccg cccctaagtc
     4381 gggaaggttc cttgcggttc gcggcgtgcc ggacgtgaca aacggaagcc gcacgtctca
     4441 ctagtaccct cgcagacgga cagcgccagg gagcaatggc agcgcgccga ccgcgatggg
     4501 ctgtggccaa tagcggctgc tcagcagggc gcgccgagag cagcggccgg gaaggggcgg
     4561 tgcgggaggc ggggtgtggg gcggtagtgt gggccctgtt cctgcccgcg cggtgttccg
     4621 cattctgcaa gcctccggag cgcacgtcgg cagtcggctc cctcgttgac cgaatcaccg
     4681 acctctctcc ccagggggat ccaccggagc ttaccatgac cgagtacaag cccacggtgc
     4741 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
     4801 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
     4861 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     4921 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     4981 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     5041 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     5101 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     5161 aggcggccga gcgcgccggg gtgcccgcct tcctggaaac ctccgcgccc cgcaacctcc
     5221 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     5281 gcacctggtg catgacccgc aagcccggtg cctgacgccc gccccacgac ccgcagcgcc
     5341 cgaccgaaag gagcgcacga ccccatgcat cggtaccttt aagaccaatg acttacaagg
     5401 cagctgtaga tcttagccac tttctagagt cggggcggcc ggccgcttcg agcagacatg
     5461 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     5521 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     5581 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     5641 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc gtcgaccgat
     5701 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
     5761 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
     5821 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5881 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5941 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6001 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6061 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6121 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6181 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6241 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6301 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6361 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6421 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     6481 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6541 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6601 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6661 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6721 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6781 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6841 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6901 gggacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6961 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7021 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7081 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7141 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7201 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7261 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7321 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7381 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7441 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7501 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7561 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7621 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7681 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7741 aagtgccacc tg
//
