LOCUS       Exported                9744 bp DNA     circular SYN 27-FEB-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9744)
  AUTHORS   Jason Vevea
  TITLE     Direct Submission
  JOURNAL   Exported Dec 1, 2021 from SnapGene Viewer 6.0.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9744
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1680..1913
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     primer_bind     2582..2624
                     /label=NC32 For IF
     promoter        2602..3048
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     primer_bind     complement(3030..3065)
                     /label=NC32 Rev IF
     CDS             3086..3139
                     /codon_start=1
                     /label=BiP signal sequence
                     /translation="MKLSLVAAMLLLLSAARA"
     CDS             3173..3271
                     /codon_start=1
                     /label=CD4 TMD
                     /translation="VLPTWSTPVQPMALIVLGGVAGLLLFIGLGIFF"
     misc_feature    3206..3271
                     /label=TMD
     CDS             3272..3307
                     /codon_start=1
                     /product="Tribasic golgi export signal KRRERRR
                     Parmar, H.B., Barry, C., Kai, F. and Duncan, R., 2014. 
                     Golgi complex?plasma membrane trafficking directed by an 
                     autonomous, tribasic Golgi export signal. Molecular biology
                     of the cell, 25(6), pp.866-878."
                     /label=Golgi Export signal
                     /translation="KYLQKRRERRRQ"
     misc_feature    3308..4390
                     /label=Syt7a Delta-TMD
     misc_feature    4436..5024
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     CDS             complement(4907..4918)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5027..5043)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             5549..5729
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    5761..5985
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6031..6459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6473..6802
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6653..6788
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6850..6897
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6916..7290
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    7420..7541
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7590..7606)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7614..7630
                     /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(7638..7668)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7683..7704
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7992..8580)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8751..9611)
                     /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(9612..9716)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta aagtgcaagt ggttttagga ccaggatgag gcggggtggg
     2641 ggtgcctacc tgacgaccga ccccgaccca ctggacaagc acccaacccc cattccccaa
     2701 attgcgcatc ccctatcaga gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca
     2761 ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc
     2821 cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc ccccgtaaac tccccttccc
     2881 ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc
     2941 gagatagggg ggcacgggcg cgaccatctg cgctgcggcg ccggcgactc agcgctgcct
     3001 cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga gagcgcaggt acggctagcg
     3061 gatccggtac cgcgggcccg ccaccatgaa gctctccctg gtggccgcga tgctgctgct
     3121 gctcagcgcg gcgcgggcca gcggtggcgg cggaagtgga ggtggaggct cggttctgcc
     3181 cacatggtcc accccggtgc agccaatggc cctgattgtg ctggggggcg tcgccggcct
     3241 cctgcttttc attgggctag gcatcttctt caagtacctg cagaaaagaa gggagcggag
     3301 acgccagcgc aaactgggca aacgctacaa gaattccttg gagacggtgg gcacgccaga
     3361 ctcggggcgt gggcgcggtg agaagaaagc catcaagttg cctgcaggag ggaaggctgt
     3421 gaatacagcc ccagtgcccg gccagacgcc acacgatgag tctgaccgca gaacggagac
     3481 ccgttcctct gtctcggacc tcgtcaactc ccttaccagc gagatgctca tgctctcccc
     3541 gggttctgag gaggatgagg cccacgaggg ctgcagccga gagaacctgg gccgaatcca
     3601 gttcagtgtt ggctacaact tccaagagtc cacactcacc gtgaaggtca tgaaggccca
     3661 agagctgcca gccaaggact tcagcggtac tagtgacccc tttgtcaaga tctacctgct
     3721 acccgacaag aagcacaaac tggagaccaa ggtgaagcgg aagaatctaa acccgcactg
     3781 gaatgagacc tttctatttg aagggtttcc ctacgagaaa gtggtgcaga gggtcctcta
     3841 cctccaggtc ctggattatg accgtttcag ccgcaatgac cccattgggg aggtgtccat
     3901 ccctctgaac aaggtggacc tgacccagat gcagaccttc tggaaggatc tgaagccatg
     3961 cagcgatggg agtgggagcc gaggggagct gctcttgtcc ctctgctaca acccctctgc
     4021 caactccatc atcgtgaaca tcatcaaagc tcgaaacctc aaagccatgg acatcggggg
     4081 cacatcagac ccctatgtga aggtgtggct gatgtataaa gacaagcggg tagagaaaaa
     4141 gaagaccgtg acaaagaaga ggaacctgaa ccccatcttc aatgagtctt tcgccttcga
     4201 catacccacg gagaagctga gggagaccac gatcatcatc actgtcatgg acaaagacaa
     4261 gctcagccgc aacgacgtca tcggcaagat ctacctgtcc tggaagagcg gaccaggtga
     4321 agtgaaacac tggaaggaca tgatcgctcg tccccggcag cctgtggccc agtggcacca
     4381 gctgaaagcc taaggccgca ctcctcagct agaattcgat atcaagctta tcgataatca
     4441 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     4501 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     4561 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     4621 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     4681 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     4741 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     4801 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     4861 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     4921 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     4981 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc
     5041 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga
     5101 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt
     5161 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     5221 ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat
     5281 ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc cagggatcag
     5341 atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga
     5401 agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga
     5461 tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat
     5521 ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg
     5581 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     5641 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     5701 cttttagtca gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga
     5761 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     5821 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     5881 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     5941 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     6001 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     6061 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     6121 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     6181 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     6241 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     6301 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     6361 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     6421 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     6481 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     6541 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     6601 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     6661 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     6721 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     6781 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     6841 tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
     6901 gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac
     6961 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag
     7021 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac
     7081 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac
     7141 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc
     7201 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc
     7261 gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc
     7321 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     7381 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     7441 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     7501 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     7561 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     7621 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     7681 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     7741 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     7801 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     7861 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     7921 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     7981 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     8041 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     8101 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     8161 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     8221 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     8281 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     8341 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     8401 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     8461 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     8521 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     8581 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     8641 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     8701 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     8761 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     8821 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     8881 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     8941 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     9001 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     9061 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     9121 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     9181 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     9241 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     9301 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     9361 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     9421 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     9481 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     9541 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     9601 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     9661 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     9721 tttccccgaa aagtgccacc tgac
//
