LOCUS       sequence-245773-        8784 bp    DNA     circular SYN 14-APR-2025
DEFINITION  lentiviral bicistronic exprression of both human alpha-1 anitrypsin 
            and ZsGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8784)
  AUTHORS   Wilson AA, Kwok LW, Hovav AH, Ohle SJ, Little FF, Fine A, Kotton DN
  TITLE     Sustained expression of alpha1-antitrypsin after transplantation of 
            manipulated hematopoietic stem cells.
  JOURNAL   Am J Respir Cell Mol Biol. 2008 Aug . 39(2):133-41.
  PUBMED    18323534
REFERENCE   2  (bases 1 to 8784)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Am J Respir
            Cell Mol Biol. 2008 Aug . 39(2):133-41."
COMMENT     Sequence Label: pHAGE-UBC-hAAT-IRES-ZsGreen-W
FEATURES             Location/Qualifiers
     source          1..8784
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2218..2616
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     misc_feature    3884..4451
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3901..4453
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4045..4062)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4272..4291
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4465..5160
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     misc_feature    5174..5762
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5227..5247)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5645..5656)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5837..6070
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6189..6207)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6275..6379
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6380..6448,6449..7240)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6598..6617)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7411..7999
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7900..7919
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8153..8170
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8410..8431
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8446..8451,8452..8469,8470..8476)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8484..8500
                     /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)."
     primer_bind     8489..8511
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8508..8524
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8508..8524
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8574..8593
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtggc
     2221 ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct cctcacggcg agcgctgcca
     2281 cgtcagacga agggcgcagc gagcgtcctg atccttccgc ccggacgctc aggacagcgg
     2341 cccgctgctc ataagactcg gccttagaac cccagtatca gcagaaggac attttaggac
     2401 gggacttggg tgactctagg gcactggttt tctttccaga gagcggaaca ggcgaggaaa
     2461 agtagtccct tctcggcgat tctgcggagg gatctccgtg gggcggtgaa cgccgatgat
     2521 tatataagga cgcgccgggt gtggcacagc tagttccgtc gcagccggga tttgggtcgc
     2581 ggttcttgtt tgtggatcgc tgtgatcgtc acttggcggc cgcatgccgt cttctgtctc
     2641 gtggggcatc ctcctgctgg caggcctgtg ctgcctggtc cctgtctccc tggctgagga
     2701 tccccaggga gatgctgccc agaagacaga tacatcccac catgatcagg atcacccaac
     2761 cttcaacaag atcaccccca acctggctga gttcgccttc agcctatacc gccagctggc
     2821 acaccagtcc aacagcacca atatcttctt ctccccagtg agcatcgcta cagcctttgc
     2881 aatgctctcc ctggggacca aggctgacac tcacgatgaa atcctggagg gcctgaattt
     2941 caacctcacg gagattccgg aggctcagat ccatgaaggc ttccaggaac tcctccgtac
     3001 cctcaaccag ccagacagcc agctccagct gaccaccggc aatggcctgt tcctcagcga
     3061 gggcctgaag ctagtggata agtttttgga ggatgttaaa aagttgtacc actcagaagc
     3121 cttcactgtc aacttcgggg acaccgaaga ggccaagaaa cagatcaacg attacgtgga
     3181 gaagggtact caagggaaaa ttgtggattt ggtcaaggag cttgacagag acacagtttt
     3241 tgctctggtg aattacatct tctttaaagg caaatgggag agaccctttg aagtcaagga
     3301 caccgaggaa gaggacttcc acgtggacca ggtgaccacc gtgaaggtgc ctatgatgaa
     3361 gcgtttaggc atgtttaaca tccagcactg taagaagctg tccagctggg tgctgctgat
     3421 gaaatacctg ggcaatgcca ccgccatctt cttcctgcct gatgagggga aactacagca
     3481 cctggaaaat gaactcaccc acgatatcat caccaagttc ctggaaaatg aagacagaag
     3541 gtctgccagc ttacatttac ccaaactgtc cattactgga acctatgatc tgaagagcgt
     3601 cctgggtcaa ctgggcatca ctaaggtctt cagcaatggg gctgacctct ccggggtcac
     3661 agaggaggca cccctgaagc tctccaaggc cgtgcataag gctgtgctga ccatcgacga
     3721 gaaagggact gaagctgctg gggccatgtt tttagaggcc atacccatgt ctatcccccc
     3781 cgaggtcaag ttcaacaaac cctttgtctt cttaatgatt gaacaaaata ccaagtctcc
     3841 cctcttcatg ggaaaagtgg tgaatcccac ccaaaaataa agatccctcc ccccccccta
     3901 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
     3961 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
     4021 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
     4081 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
     4141 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     4201 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     4261 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     4321 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
     4381 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     4441 cacgatgata atatggccac acatatggcc cagtccaagc acggcctgac caaggagatg
     4501 accatgaagt accgcatgga gggctgcgtg gacggccaca agttcgtgat caccggcgag
     4561 ggcatcggct accccttcaa gggcaagcag gccatcaacc tgtgcgtggt ggagggcggc
     4621 cccttgccct tcgccgagga catcttgtcc gccgccttca tgtacggcaa ccgcgtgttc
     4681 accgagtacc cccaggacat cgtcgactac ttcaagaact cctgccccgc cggctacacc
     4741 tgggaccgct ccttcctgtt cgaggacggc gccgtgtgca tctgcaacgc cgacatcacc
     4801 gtgagcgtgg aggagaactg catgtaccac gagtccaagt tctacggcgt gaacttcccc
     4861 gccgacggcc ccgtgatgaa gaagatgacc gacaactggg agccctcctg cgagaagatc
     4921 atccccgtgc ccaagcaggg catcttgaag ggcgacgtga gcatgtacct gctgctgaag
     4981 gacggtggcc gcttgcgctg ccagttcgac accgtgtaca aggccaagtc cgtgccccgc
     5041 aagatgcccg actggcactt catccagcac aagctgaccc gcgaggaccg cagcgacgcc
     5101 aagaaccaga agtggcacct gaccgagcac gccatcgcct ccggctccgc cttgccctga
     5161 atcgatagat cctaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     5221 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     5281 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     5341 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     5401 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     5461 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     5521 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     5581 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     5641 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     5701 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     5761 gcctgagatc ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     5821 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt
     5881 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     5941 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     6001 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     6061 atctctagca gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa
     6121 tgaatatcag agagtgagag gcccgggtta attaaggaaa gggctagatc attcttgaag
     6181 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     6241 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     6301 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     6361 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     6421 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     6481 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     6541 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     6601 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     6661 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     6721 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     6781 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     6841 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     6901 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     6961 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     7021 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     7081 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     7141 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     7201 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     7261 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     7321 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     7381 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     7441 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     7501 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     7561 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     7621 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     7681 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     7741 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     7801 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     7861 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     7921 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     7981 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     8041 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     8101 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     8161 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     8221 gattcattaa tgcagcaagc tcatggctga ctaatttttt ttatttatgc agaggccgag
     8281 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     8341 ttttgcaaaa agctccccgt ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     8401 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     8461 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     8521 tgacatgatt acgaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     8581 gtccaaactc atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca
     8641 aaatcatccc aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca
     8701 tttactctaa acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta
     8761 aatatgtact acaaacttag tagt
//
