LOCUS       62056_14950        8714 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8714)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..507
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             522..2171
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     LTR             2245..2478
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2550..2684
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      2711..2846
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(2867..2885)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2895..2911)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3053..3508
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3534..3638
                     /label=AmpR promoter
     CDS             3639..4496
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4670..5258
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5546..5567
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5582..5612
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5620..5636
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5644..5660
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5681..5699
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5727..5953
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     misc_feature    6181..6306
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    6799..7032
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7217..7261
                     /codon_start=1
                     /label=gp41 peptide
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et 
                     al., 2013)"
                     /translation="KNEQELLELDKWASL"
     primer_bind     7619..7635
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             7787..8503
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    8545..8662
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        8711..8714
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
ORIGIN
        1 ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg
       61 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt
      121 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct
      181 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa
      241 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc
      301 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc
      361 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc
      421 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt
      481 gaccgaatca ccgacctctc tccccagggg gatccgccac catggaagac gccaaaaaca
      541 taaagaaagg cccggcgcca ttctatcctc ttgaggatgg aaccgctgga gagcaactgc
      601 ataaggctat gaagagatac gccctggttc ctggaacaat tgcttttaca gatgcacata
      661 tcgaggtgaa catcacgtac gcggaatact tcgaaatgtc cgttcggttg gcagaagcta
      721 tgaaacgata tgggctgaat acaaatcaca gaatcgtcgt atgcagtgaa aactctcttc
      781 aattctttat gccggtgttg ggcgcgttat ttatcggagt tgcagttgcg cccgcgaacg
      841 acatttataa tgaacgtgaa ttgctcaaca gtatgaacat ttcgcagcct accgtagtgt
      901 ttgtttccaa aaaggggttg caaaaaattt tgaacgtgca aaaaaaatta ccaataatcc
      961 agaaaattat tatcatggat tctaaaacgg attaccaggg atttcagtcg atgtacacgt
     1021 tcgtcacatc tcatctacct cccggtttta atgaatacga ttttgtacca gagtcctttg
     1081 atcgtgacaa aacaattgca ctgataatga actcctctgg atctactggg ttacctaagg
     1141 gtgtggccct tccgcataga actgcctgcg tcagattctc gcatgccaga gatcctattt
     1201 ttggcaatca aatcattccg gatactgcga ttttaagtgt tgttccattc catcacggtt
     1261 ttggaatgtt tactacactc ggatatttga tatgtggatt tcgagtcgtc ttaatgtata
     1321 gatttgaaga agagctgttt ttacgatccc ttcaggatta caaaattcaa agtgcgttgc
     1381 tagtaccaac cctattttca ttcttcgcca aaagcactct gattgacaaa tacgatttat
     1441 ctaatttaca cgaaattgct tctgggggcg cacctctttc gaaagaagtc ggggaagcgg
     1501 ttgcaaaacg cttccatctt ccagggatac gacaaggata tgggctcact gagactacat
     1561 cagctattct gattacaccc gagggggatg ataaaccggg cgcggtcggt aaagttgttc
     1621 cattttttga agcgaaggtt gtggatctgg ataccgggaa aacgctgggc gttaatcaga
     1681 gaggcgaatt atgtgtcaga ggacctatga ttatgtccgg ttatgtaaac aatccggaag
     1741 cgaccaacgc cttgattgac aaggatggat ggctacattc tggagacata gcttactggg
     1801 acgaagacga acacttcttc atagttgacc gcttgaagtc tttaattaaa tacaaaggat
     1861 accaggtggc ccccgctgaa ttggagtcga tattgttaca acaccccaac atcttcgacg
     1921 cgggcgtggc aggtcttccc gacgatgacg ccggtgaact tcccgccgcc gttgttgttt
     1981 tggagcacgg aaagacgatg acggaaaaag agatcgtgga ttacgtcgcc agtcaagtaa
     2041 caaccgcgaa aaagttgcgc ggaggagttg tgtttgtgga cgaagtaccg aaaggtctta
     2101 ccggaaaact cgacgcaaga aaaatcagag agatcctcat aaaggccaag aagggcggaa
     2161 agtccaaatt gtaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     2221 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     2281 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     2341 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     2401 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     2461 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     2521 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
     2581 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     2641 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
     2701 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     2761 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     2821 gtagtgagga ggcttttttg gaggcctagg gacgtaccca attcgcccta tagtgagtcg
     2881 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     2941 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     3001 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc
     3061 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3121 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3181 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3241 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3301 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3361 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3421 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3481 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     3541 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3601 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3661 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3721 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     3781 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     3841 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     3901 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     3961 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4021 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4081 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     4141 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     4201 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4261 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     4321 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     4381 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     4441 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4501 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4561 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4621 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4681 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4741 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4801 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4861 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4921 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4981 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5041 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5101 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5161 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5221 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5281 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     5341 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     5401 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     5461 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     5521 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     5581 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     5641 acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg
     5701 aacaaaagct ggagctgcaa gcttaatgta gtcttatgca atactcttgt agtcttgcaa
     5761 catggtaacg atgagttagc aacatgcctt acaaggagag aaaaagcacc gtgcatgccg
     5821 attggtggaa gtaaggtggt acgatcgtgc cttattagga aggcaacaga cgggtctgac
     5881 atggattgga cgaaccactg aattgccgca ttgcagagat attgtattta agtgcctagc
     5941 tcgatacata aacgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     6001 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     6061 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     6121 gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag
     6181 ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga
     6241 ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag
     6301 agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca
     6361 gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga
     6421 ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag
     6481 ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca
     6541 accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag
     6601 atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag
     6661 acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt
     6721 aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga
     6781 aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac
     6841 tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt
     6901 gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac
     6961 agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga
     7021 tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc
     7081 ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat
     7141 ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc
     7201 gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt
     7261 gtggaattgg tttaacataa caaattggct gtggtatata aaattattca taatgatagt
     7321 aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag
     7381 gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg ctagcccggc
     7441 cagtgccaag ttgagacaca ttccacacat tccactgcaa gcttgagaca cattccacac
     7501 attccactgc aagcttctag cccggccagt gccaagttga gacacattcc acacattcca
     7561 ctgcaagctt gagacacatt ccacacattc cactgcaagc ttctagagat ctgcaggtcg
     7621 aggtcgacgg tatcgataag cttgggggtg ggcgccgggg ggaccttaaa gcctctgccc
     7681 cccaaggagc ccttcccaga cagccgccgg cacccaccgc tccgtgggac gatcccccaa
     7741 gagcttggca ttccggtact gttggtaaag cgacgtcgac gccaccatgg tgagcaaggg
     7801 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     7861 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     7921 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     7981 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     8041 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     8101 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     8161 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     8221 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     8281 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     8341 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     8401 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     8461 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaattct cgacctcgag
     8521 acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc
     8581 aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca
     8641 aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc cactttggcc
     8701 gcggctcgag gggg
//
