LOCUS       Exported                7152 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 7152)
  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..7152
                     /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..3055
                     /label=G1333N
     CDS             3068..3316
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     polyA_signal    3353..3577
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    3567..3581
                     /label=HR
     promoter        3584..4094
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             4116..4715
                     /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    4862..4983
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(5402..5990)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6161..7021)
                     /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(7022..7126)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7142..7152
                     /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 agtaatgggg gtggcaaaca ggctcttgag actgttcagc gccttctacc
     1381 agttctctgt caagcccacg gcctgacgcc cgagcaagtt gtagcgattg ctagtaatat
     1441 tggtggcaaa caggctcttg agacggttca gcgcctcctt ccagttctct gtcaagccca
     1501 cggactcacc ccagatcagg ttgtagcgat tgctagtaac aatggtggca aacaggctct
     1561 tgaaaccgta cagcgcctac tgccagttct ctgtcaagcc cacggtctga ctccggagca
     1621 agttgtagcg attgctagta acaatggtgg caaacaggct cttgaaaccg tacagcgcct
     1681 actgccagtt ctctgtcaag cccacggtct gactccggag caagtggtag cgattgctag
     1741 taacaatggt ggcaaacagg ctcttgaaac cgtacagcgc ctactgccag ttctctgtca
     1801 agcccacggt ctgactccgg agcaagttgt agcgattgct agtaatattg gtggcaaaca
     1861 ggctcttgag acggttcagc gcctccttcc agttctctgt caagcccacg gactcacccc
     1921 agatcaagtt gtagcgattg ctagtaatgg gggtggcaaa caggctcttg agactgttca
     1981 gcgccttcta ccagttctct gtcaagccca cggcctgacg cccgagcaag ttgtagcgat
     2041 tgctagtaat attggtggca aacaggctct tgagacggtt cagcgcctcc ttccagttct
     2101 ctgtcaagcc cacggactca ccccagatca 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 atattggtgg caaacaggct cttgagacgg ttcagcgcct
     2701 ccttccagtt ctctgtcaag cccacggact caccccagat caagttgtag cgattgcaag
     2761 taatgggggc ggcaggccgg cgctggagag cattgttgcc cagttatctc gccctgatcc
     2821 ggcgttggcc gcgttgacca acgaccacct cgtcgccttg gcctgcctcg gcggacgtcc
     2881 tgcgctggat gcagtgaaaa agggattggg aggctccgct agcggctcct acgccctggg
     2941 cccatatcag atctctgccc cacagctgcc agcatacaac ggacagaccg tgggcacatt
     3001 ctactatgtg aatgacgccg gcggcctgga gtctaaggtg tttagctccg gcggctctgg
     3061 aggcagcacc aacctgtctg acatcatcga gaaggagaca ggcaagcagc tggtcatcca
     3121 ggagagcatc ctgatgctgc ccgaagaagt cgaagaagtg atcggaaaca agcctgagag
     3181 cgatatcctg gtccataccg cctacgacga gagtaccgac gaaaatgtga tgctgctgac
     3241 atccgacgcc ccagagtata agccctgggc tctggtcatc caggattcca acggagagaa
     3301 caaaatcaaa atgctgctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     3361 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3421 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3481 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3541 aatagcaggc atgctgggga tgcggtgggc tctatggctc gagggggttg gggttgcgcc
     3601 ttttccaagg cagccctggg tttgcgcagg gacgcggctg ctctgggcgt ggttccggga
     3661 aacgcagcgg cgccgaccct gggactcgca cattcttcac gtccgttcgc agcgtcaccc
     3721 ggatcttcgc cgctaccctt gtgggccccc cggcgacgct tcctgctccg cccctaagtc
     3781 gggaaggttc cttgcggttc gcggcgtgcc ggacgtgaca aacggaagcc gcacgtctca
     3841 ctagtaccct cgcagacgga cagcgccagg gagcaatggc agcgcgccga ccgcgatggg
     3901 ctgtggccaa tagcggctgc tcagcagggc gcgccgagag cagcggccgg gaaggggcgg
     3961 tgcgggaggc ggggtgtggg gcggtagtgt gggccctgtt cctgcccgcg cggtgttccg
     4021 cattctgcaa gcctccggag cgcacgtcgg cagtcggctc cctcgttgac cgaatcaccg
     4081 acctctctcc ccagggggat ccaccggagc ttaccatgac cgagtacaag cccacggtgc
     4141 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
     4201 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
     4261 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     4321 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     4381 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     4441 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     4501 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     4561 aggcggccga gcgcgccggg gtgcccgcct tcctggaaac ctccgcgccc cgcaacctcc
     4621 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     4681 gcacctggtg catgacccgc aagcccggtg cctgacgccc gccccacgac ccgcagcgcc
     4741 cgaccgaaag gagcgcacga ccccatgcat cggtaccttt aagaccaatg acttacaagg
     4801 cagctgtaga tcttagccac tttctagagt cggggcggcc ggccgcttcg agcagacatg
     4861 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     4921 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     4981 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     5041 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc gtcgaccgat
     5101 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
     5161 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
     5221 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5281 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5341 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5401 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5461 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5521 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5581 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5641 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5701 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5761 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5821 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     5881 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5941 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6001 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6061 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6121 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6181 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6241 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6301 gggacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6361 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6421 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6481 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6541 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6601 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6661 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6721 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6781 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6841 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6901 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6961 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7021 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7081 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7141 aagtgccacc tg
//
