LOCUS       Exported                9316 bp ds-DNA     circular SYN 05-MAR-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9316)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Mar 6, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9316
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        310..689
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             907..1087
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1134..1259
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1758..1991
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2518..2635
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        3174..3477
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3478..3681
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    3718..4056
                     /label=MCP
     misc_feature    4069..4410
                     /label=MCP
     CDS             4435..4986
                     /codon_start=1
                     /label=P65
                     /translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP
                     GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE
                     FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED
                     FSSIADMDFSALLSQISSSGQ"
     CDS             5002..5373
                     /codon_start=1
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     CDS             5389..5442
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             5443..6159
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     protein_bind    6179..6212
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    6268..6856
                     /label=WPRE
                     /note="woodchuck posttranscriptional regulatory element"
     CDS             complement(6739..6750)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6859..6875)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             7385..7565
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7627..8215)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8386..9246)
                     /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(join(9247..9316,1..35))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
       61 gacgcttaag cggtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
      121 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
      181 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
      241 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
      301 atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
      361 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      421 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      481 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      541 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      601 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      661 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      721 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      781 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      841 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcagc gcgttttgcc
      901 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
      961 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     1021 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     1081 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
     1141 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
     1201 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
     1261 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
     1321 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
     1381 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
     1441 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
     1501 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
     1561 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cggccgcgct gatcttcaga
     1621 cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta
     1681 aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa
     1741 aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact
     1801 atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg
     1861 cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca
     1921 gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat
     1981 caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct
     2041 tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg
     2101 gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg
     2161 caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg
     2221 tggaattggt ttaacataac aaattggctg tggtatataa aattattcat aatgatagta
     2281 ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg
     2341 cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg
     2401 cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg
     2461 aacggatcgg cactgcgtgc gccaattctg cagacaaatg gcagtattca tccacaattt
     2521 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     2581 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt
     2641 ttattacagg gacagcagag atccagtttg gttagtaccg ggcccgctct agacgctatg
     2701 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2761 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     2821 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     2881 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     2941 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3001 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgctcg
     3061 agtactagga tccattaggc ggccgcgtgg ataaccgtat taccgccatg cattagttat
     3121 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     3181 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     3241 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     3301 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     3361 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     3421 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     3481 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     3541 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     3601 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     3661 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcc caccatggct
     3721 tctaacttta ctcagttcgt tctcgtcgac aatggcggaa ctggcgacgt gactgtcgcc
     3781 ccaagcaact tcgctaacgg gatcgctgaa tggatcagct ctaactcgcg ttcacaggct
     3841 tacaaagtaa cctgtagcgt tcgtcagagc tctgcgcaga atcgcaaata caccatcaaa
     3901 gtcgaggtgc ctaaaggcgc ctggcgttcg tacttaaata tggaactaac cattccaatt
     3961 ttcgccacga attccgactg cgagcttatt gttaaggcaa tgcaaggtct cctaaaagat
     4021 ggaaacccga ttccctcagc aatcgcagca aactccggca tctacgccat ggccagcaac
     4081 ttcacccagt tcgtgctggt ggacaacggc ggcaccggcg acgtgaccgt ggcccccagc
     4141 aacttcgcca acggcatcgc cgagtggatc agcagcaaca gcagaagcca ggcctacaag
     4201 gtgacctgca gcgtgagaca gagcagcgcc cagaacagaa agtacaccat caaggtggag
     4261 gtgcccaagg gcgcctggag aagctacctg aacatggagc tgaccatccc catcttcgcc
     4321 accaacagcg actgcgagct gatcgtgaag gccatgcagg gcctgctgaa ggacggcaac
     4381 cccatcccca gcgccatcgc cgccaacagc ggcatctacg ccgacccacc ggtcccttca
     4441 gggcagatca gcaaccaggc cctggctctg gcccctagct ccgctccagt gctggcccag
     4501 actatggtgc cctctagtgc tatggtgcct ctggcccagc cacctgctcc agcccctgtg
     4561 ctgaccccag gaccacccca gtcactgagc gctccagtgc ccaagtctac acaggccggc
     4621 gaggggactc tgagtgaagc tctgctgcac ctgcagttcg acgctgatga ggacctggga
     4681 gctctgctgg ggaacagcac cgatcccgga gtgttcacag atctggcctc cgtggacaac
     4741 tctgagtttc agcagctgct gaatcagggc gtgtccatgt ctcatagtac agccgaacca
     4801 atgctgatgg agtaccccga agccattacc cggctggtga ccggcagcca gcggcccccc
     4861 gaccccgctc caactcccct gggaaccagc ggcctgccta atgggctgtc cggagatgaa
     4921 gacttctcaa gcatcgctga tatggacttt agtgccctgc tgtcacagat ttcctctagt
     4981 gggcagggag gaggtggaag cggcttcagc gtggacacca gtgccctgct ggacctgttc
     5041 agcccctcgg tgaccgtgcc cgacatgagc ctgcctgacc ttgacagcag cctggccagt
     5101 atccaagagc tcctgtctcc ccaggagccc cccaggcctc ccgaggcaga gaacagcagc
     5161 ccggattcag ggaagcagct ggtgcactac acagcgcagc cgctgttcct gctggacccc
     5221 ggctccgtgg acaccgggag caacgacctg ccggtgctgt ttgagctggg agagggctcc
     5281 tacttctccg aaggggacgg cttcgccgag gaccccacca tctccctgct gacaggctcg
     5341 gagcctccca aagccaagga ccccactgtc tccgctagcg gcagtggaga gggcagagga
     5401 agtctgctaa catgcggtga cgtcgaggag aatcctggcc cagtgagcaa gggcgaggag
     5461 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     5521 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     5581 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     5641 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     5701 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     5761 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     5821 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     5881 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     5941 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     6001 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     6061 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     6121 gccgggatca ctctcggcat ggacgagctg tacaagtaag aattcgtcga gggacctaat
     6181 aacttcgtat agcatacatt atacgaagtt atacatgttt aagggttccg gttccactag
     6241 gtacaattcg atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga
     6301 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     6361 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     6421 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     6481 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     6541 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     6601 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     6661 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     6721 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     6781 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     6841 tgggccgcct ccccgcatcg ataccgtcga cctcgatcga gacctagaaa aacatggagc
     6901 aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga
     6961 ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa
     7021 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     7081 ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag gctacttccc
     7141 tgattggcag aactacacac cagggccagg gatcagatat ccactgacct ttggatggtg
     7201 ctacaagcta gtaccagttg agcaagagaa ggtagaagaa gccaatgaag gagagaacac
     7261 ccgcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag aagtattaga
     7321 gtggaggttt gacagccgcc tagcatttca tcacatggcc cgagagctgc atccggactg
     7381 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     7441 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     7501 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     7561 tagcagcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     7621 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     7681 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     7741 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7801 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7861 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     7921 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7981 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     8041 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     8101 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     8161 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     8221 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     8281 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     8341 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     8401 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     8461 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     8521 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     8581 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     8641 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     8701 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     8761 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     8821 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     8881 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     8941 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     9001 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     9061 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     9121 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     9181 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     9241 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     9301 cggatacata tttgaa
//
