LOCUS       Exported                6796 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pHCMV-AmphoEnv
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6796)
  AUTHORS   Sena-Esteves M, Tebbets JC, Steffens S, Crombleholme T, Flake AW
  TITLE     Optimized large-scale production of high titer lentivirus vector 
            pseudotypes.
  JOURNAL   J Virol Methods. 2004 Dec 15. 122(2):131-9.
  PUBMED    15542136
REFERENCE   2  (bases 1 to 6796)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6796
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        97..476
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        477..680
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     631..651
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     677..701
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          792..1364
                     /label=-beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             1441..3405
                     /codon_start=1
                     /gene="env"
                     /product="murine leukemia virus gPr80 envelope polyprotein 
                     precursor"
                     /label=env
                     /note="proteolytically cleaved into gp70SU and p15TM 
                     subunits"
                     /translation="MARSTLSKPPQDKINPWKPLIVMGVLLGVGMAESPHQVFNVTWRV
                     TNLMTGRTANATSLLGTVQDAFPKLYFDLCDLVGEEWDPSDQEPYVGYGCKYPAGRQRT
                     RTFDFYVCPGHTVKSGCGGPGEGYCGKWGCETTGQAYWKPTSSWDLISLKRGNTPWDTG
                     CSKVACGPCYDLSKVSNSFQGATRGGRCNPLVLEFTDAGKKANWDGPKSWGLRLYRTGT
                     DPITMFSLTRQVLNVGPRVPIGPNPVLPDQRLPSSPIEIVPAPQPPSPLNTSYPPSTTS
                     TPSTSPTSPSVPQPPPGTGDRLLALVKGAYQALNLTNPDKTQECWLCLVSGPPYYEGVA
                     VVGTYTNHSTAPANCTATSQHKLTLSEVTGQGLCMGAVPKTHQALCNTTQSAGSGSYYL
                     AAPAGTMWACSTGLTPCLSTTVLNLTTDYCVLVELWPRVIYHSPDYMYGQLEQRTKYKR
                     EPVSLTLALLLGGLTMGGIAAGIGTGTTALIKTQQFEQLHAAIQTDLNEVEKSITNLEK
                     SLTSLSEVVLQNRRGLDLLFLKEGGLCAALKEECCFYADHTGLVRDSMAKLRERLNQRQ
                     KLFETGQGWFEGLFNRSPWFTTLISTIMGPLIVLLLILLFGPCILNRLVQFVKDRISVV
                     QALVLTQQYHQLKPIEYEP"
     RBS             2436..2444
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(3500..3519)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3565..3620
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3619..3638)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3978..3994)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3978..3994)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3991..4013)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4002..4018
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     promoter        complement(4026..4056)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4071..4092
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4209..4226)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4380..4968)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4460..4479)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5437..5816)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     5763..5782
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6001..6105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6131..6586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6268..6289)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6480..6499
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6713..6735
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6727..6744
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6728..6744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6754..6773
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6754..6772
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gcggccgctc tagagagctt ggcccattgc atacgttgta tccatatcat aatatgtaca
       61 tttatattgg ctcatgtcca acattaccgc catgttgaca ttgattattg actagttatt
      121 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      181 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      241 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      301 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      361 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      421 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      481 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      541 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      601 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      661 aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac
      721 gctgttttga cctccataga agacaccggg accgatccag cctccggtcg accgatcctg
      781 agaacttcag ggtgagtttg gggacccttg attgttcttt ctttttcgct attgtaaaat
      841 tcatgttata tggagggggc aaagttttca gggtgttgtt tagaatggga agatgtccct
      901 tgtatcacca tggaccctca tgataatttt gtttctttca ctttctactc tgttgacaac
      961 cattgtctcc tcttattttc ttttcatttt ctgtaacttt ttcgttaaac tttagcttgc
     1021 atttgtaacg aatttttaaa ttcacttttg tttatttgtc agattgtaag tactttctct
     1081 aatcactttt ttttcaaggc aatcagggta tattatattg tacttcagca cagttttaga
     1141 gaacaattgt tataattaaa tgataaggta gaatatttct gcatataaat tctggctggc
     1201 gtggaaatat tcttattggt agaaacaact acaccctggt catcatcctg cctttctctt
     1261 tatggttaca atgatataca ctgtttgaga tgaggataaa atactctgag tccaaaccgg
     1321 gcccctctgc taaccatgtt catgccttct tctctttcct acagctcctg ggcaacgtgc
     1381 tggttgttgt gctgtctcat cattttggca aagaattcct cgacggatct gggccgcacc
     1441 atggcgcgtt caacgctctc aaaaccccct caagataaga ttaacccgtg gaagccctta
     1501 atagtcatgg gagtcctgtt aggagtaggg atggcagaga gcccccatca ggtctttaat
     1561 gtaacctgga gagtcaccaa cctgatgact gggcgtaccg ccaatgccac ctccctcctg
     1621 ggaactgtac aagatgcctt cccaaaatta tattttgatc tatgtgatct ggtcggagag
     1681 gagtgggacc cttcagacca ggaaccgtat gtcgggtatg gctgcaagta ccccgcaggg
     1741 agacagcgga cccggacttt tgacttttac gtgtgccctg ggcataccgt aaagtcgggg
     1801 tgtgggggac caggagaggg ctactgtggt aaatgggggt gtgaaaccac cggacaggct
     1861 tactggaagc ccacatcatc gtgggaccta atctccctta agcgcggtaa caccccctgg
     1921 gacacgggat gctctaaagt tgcctgtggc ccctgctacg acctctccaa agtatccaat
     1981 tccttccaag gggctactcg agggggcaga tgcaaccctc tagtcctaga attcactgat
     2041 gcaggaaaaa aggctaactg ggacgggccc aaatcgtggg gactgagact gtaccggaca
     2101 ggaacagatc ctattaccat gttctccctg acccggcagg tccttaatgt gggaccccga
     2161 gtccccatag ggcccaaccc agtattaccc gaccaaagac tcccttcctc accaatagag
     2221 attgtaccgg ctccacagcc acctagcccc ctcaatacca gttacccccc ttccactacc
     2281 agtacaccct caacctcccc tacaagtcca agtgtcccac agccaccccc aggaactgga
     2341 gatagactac tagctctagt caaaggagcc tatcaggcgc ttaacctcac caatcccgac
     2401 aagacccaag aatgttggct gtgcttagtg tcgggacctc cttattacga aggagtagcg
     2461 gtcgtgggca cttataccaa tcattccacc gctccggcca actgtacggc cacttcccaa
     2521 cataagctta ccctatctga agtgacagga cagggcctat gcatgggggc agtacctaaa
     2581 actcaccagg ccttatgtaa caccacccaa agcgccggct caggatccta ctaccttgca
     2641 gcacccgccg gaacaatgtg ggcttgcagc actggattga ctccctgctt gtccaccacg
     2701 gtgctcaatc taaccacaga ttattgtgta ttagttgaac tctggcccag agtaatttac
     2761 cactcccccg attatatgta tggtcagctt gaacagcgta ccaaatataa aagagagcca
     2821 gtatcattga ccctggccct tctactagga ggattaacca tgggagggat tgcagctgga
     2881 atagggacgg ggaccactgc cttaattaaa acccagcagt ttgagcagct tcatgccgct
     2941 atccagacag acctcaacga agtcgaaaag tcaattacca acctagaaaa gtcactgacc
     3001 tcgttgtctg aagtagtcct acagaaccgc agaggcctag atttgctatt cctaaaggag
     3061 ggaggtctct gcgcagccct aaaagaagaa tgttgttttt atgcagacca cacggggcta
     3121 gtgagagaca gcatggccaa attaagagaa aggcttaatc agagacaaaa actatttgag
     3181 acaggccaag gatggttcga agggctgttt aatagatccc cctggtttac caccttaatc
     3241 tccaccatca tgggacctct aatagtactc ttactgatct tactctttgg accttgcatt
     3301 ctcaatcgat tagtccaatt tgttaaagac aggatatcag tggtccaggc tctagttttg
     3361 actcaacaat atcaccagct gaagcccata gagtacgagc catgataacc gtcgaggaat
     3421 tcactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg
     3481 ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc
     3541 cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt
     3601 ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc
     3661 agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca
     3721 aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat
     3781 tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt
     3841 tttctttaac atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct
     3901 cctgactact cccagtcata gctgtccctc ttctcttatg gagatccctc gacggatcgg
     3961 ccgcaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4021 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4081 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4141 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4201 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4261 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4321 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4381 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4441 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4501 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4561 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4621 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4681 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4741 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4801 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4861 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4921 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4981 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5041 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5101 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5161 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5221 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5281 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5341 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5401 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5461 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5521 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg gttagctcct tcggtcctcc
     5581 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5641 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5701 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5761 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5821 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5881 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5941 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6001 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6061 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     6181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     6241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     6301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     6361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     6421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     6481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     6541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
     6601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     6661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     6721 cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata gggcgaattg
     6781 gagctccacc gcggtg
//
