LOCUS       Exported                9361 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 9361)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Mar 6, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9361
                     /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    3715..3717
                     /label=ATG
     misc_feature    3718..4086
                     /label=PCP
     misc_feature    4096..4464
                     /label=PCP
     CDS             4480..5031
                     /codon_start=1
                     /label=P65
                     /translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP
                     GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE
                     FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED
                     FSSIADMDFSALLSQISSSGQ"
     CDS             5047..5418
                     /codon_start=1
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     CDS             5434..5487
                     /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             5488..6204
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     protein_bind    6224..6257
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    6313..6901
                     /label=WPRE
                     /note="woodchuck posttranscriptional regulatory element"
     CDS             complement(6784..6795)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6904..6920)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             7430..7610
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7672..8260)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8431..9291)
                     /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(9292..9361,1..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 caccatgttg
     3721 gcttccaaaa ctatcgttct gagcgtaggt gaagcgacca gaacgttaac tgaaattcaa
     3781 tctacagcag acaggcaaat atttgaagag aaagtcggtc cattagtagg gaggttaaga
     3841 ttgacagcta gcttaagaca aaatggagct aaaactgcat acagggtcaa tttaaaattg
     3901 gatcaagcag atgtcgttga tagcggtcta cctaaggtta gatacacgca agtgtggtca
     3961 cacgatgtca ctattgtcgc gaattctacc gaagctagta gaaaatcatt gtatgattta
     4021 acaaaatcct tggtggctac ctctcaggtg gaagatcttg ttgtgaattt agttccatta
     4081 ggtagggcgg atccgctagc ctccaaaacc atcgttcttt cggtcggcga ggctactcgc
     4141 actctgactg agatccagtc caccgcagac cgtcagatct tcgaagagaa ggtcgggcct
     4201 ctggtgggtc ggctgcgcct cacggcttcg ctccgtcaaa acggagccaa gaccgcgtat
     4261 cgcgtcaacc taaaactgga tcaggcggac gtcgttgatt ccggacttcc gaaagtgcgc
     4321 tacactcagg tatggtcgca cgacgtgaca atcgttgcga atagcaccga ggcctcgcgc
     4381 aaatcgttgt acgatttgac caagtccctc gtcgcgacct cgcaggtcga agatcttgtc
     4441 gtcaaccttg tgccgctggg ccgtccaccg gtcgccaccc cttcagggca gatcagcaac
     4501 caggccctgg ctctggcccc tagctccgct ccagtgctgg cccagactat ggtgccctct
     4561 agtgctatgg tgcctctggc ccagccacct gctccagccc ctgtgctgac cccaggacca
     4621 ccccagtcac tgagcgctcc agtgcccaag tctacacagg ccggcgaggg gactctgagt
     4681 gaagctctgc tgcacctgca gttcgacgct gatgaggacc tgggagctct gctggggaac
     4741 agcaccgatc ccggagtgtt cacagatctg gcctccgtgg acaactctga gtttcagcag
     4801 ctgctgaatc agggcgtgtc catgtctcat agtacagccg aaccaatgct gatggagtac
     4861 cccgaagcca ttacccggct ggtgaccggc agccagcggc cccccgaccc cgctccaact
     4921 cccctgggaa ccagcggcct gcctaatggg ctgtccggag atgaagactt ctcaagcatc
     4981 gctgatatgg actttagtgc cctgctgtca cagatttcct ctagtgggca gggaggaggt
     5041 ggaagcggct tcagcgtgga caccagtgcc ctgctggacc tgttcagccc ctcggtgacc
     5101 gtgcccgaca tgagcctgcc tgaccttgac agcagcctgg ccagtatcca agagctcctg
     5161 tctccccagg agccccccag gcctcccgag gcagagaaca gcagcccgga ttcagggaag
     5221 cagctggtgc actacacagc gcagccgctg ttcctgctgg accccggctc cgtggacacc
     5281 gggagcaacg acctgccggt gctgtttgag ctgggagagg gctcctactt ctccgaaggg
     5341 gacggcttcg ccgaggaccc caccatctcc ctgctgacag gctcggagcc tcccaaagcc
     5401 aaggacccca ctgtctccgc tagcggcagt ggagagggca gaggaagtct gctaacatgc
     5461 ggtgacgtcg aggagaatcc tggcccagtg agcaagggcg aggagctgtt caccggggtg
     5521 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     5581 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     5641 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     5701 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     5761 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     5821 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     5881 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     5941 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     6001 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     6061 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     6121 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     6181 ggcatggacg agctgtacaa gtaagaattc gtcgagggac ctaataactt cgtatagcat
     6241 acattatacg aagttataca tgtttaaggg ttccggttcc actaggtaca attcgatatc
     6301 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     6361 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     6421 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     6481 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     6541 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     6601 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     6661 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     6721 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     6781 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     6841 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     6901 catcgatacc gtcgacctcg atcgagacct agaaaaacat ggagcaatca caagtagcaa
     6961 tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg
     7021 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct
     7081 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     7141 agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt ggcagaacta
     7201 cacaccaggg ccagggatca gatatccact gacctttgga tggtgctaca agctagtacc
     7261 agttgagcaa gagaaggtag aagaagccaa tgaaggagag aacacccgct tgttacaccc
     7321 tgtgagcctg catgggatgg atgacccgga gagagaagta ttagagtgga ggtttgacag
     7381 ccgcctagca tttcatcaca tggcccgaga gctgcatccg gactgtactg ggtctctctg
     7441 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     7501 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     7561 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gcatgtgagc
     7621 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     7681 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     7741 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     7801 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     7861 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7921 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7981 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     8041 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     8101 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     8161 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     8221 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     8281 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     8341 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     8401 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     8461 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     8521 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     8581 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     8641 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     8701 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     8761 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     8821 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     8881 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     8941 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     9001 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     9061 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     9121 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     9181 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     9241 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     9301 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     9361 a
//
