LOCUS       sequence-204675-        7589 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty plasmid, pSIN-EF1-alpha-IRES-puro.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7589)
  AUTHORS   Kyriakakis P, Catanho M, Hoffner N, Thavarajah W, Hu VJ, Chao SS, 
            Hsu A, Pham V, Naghavian L, Dozier LE, Patrick GN, Coleman TP
  TITLE     Biosynthesis of Orthogonal Molecules Using Ferredoxin and 
            Ferredoxin-NADP(+) Reductase Systems Enables Genetically Encoded 
            PhyB Optogenetics.
  JOURNAL   ACS Synth Biol. 2018 Feb 16;7(2):706-717. doi: 
            10.1021/acssynbio.7b00413. Epub 2018 Jan 24.
  PUBMED    29301067
REFERENCE   2  (bases 1 to 7589)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.7b00413"; journalName: "ACS Synth Biol"; date: 
            "2018-02-16- 16"; volume: "7"; issue: "2"; pages: "706-717"
COMMENT     Sequence Label: pPKm-145
FEATURES             Location/Qualifiers
     source          1..7589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..1180
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          233..1171
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1128..1148
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    join(1297..1871,1872..1874,1875..1883)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1464..1481)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1691..1710
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1903..2502
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1903..1922)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2399..2419
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2705..2938
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(3089..3106)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3095..3319
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      3362..3371
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        3415..3519
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3520..3588,3589..4380)
                     /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"
     primer_bind     complement(3738..3757)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4551..5139
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5040..5059
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             5298..5931
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    5978..6103
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6596..6829
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7014..7058
                     /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    7345..7462
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
       61 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      121 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      181 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      241 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      301 tacttccacc tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg
      361 ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg
      421 gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc
      481 tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg
      541 gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc
      601 cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg
      661 agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc
      721 tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc
      781 agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag
      841 gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc
      901 gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga
      961 ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat
     1021 ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta
     1081 attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac
     1141 agtggttcaa agtttttttc ttccatttca ggtgtcgtga ggaattgatc cttcgaacta
     1201 gaattcgaag cttgagctcg agatctgagt ccggtagcgc tagttcgaag gatccgcatg
     1261 catctagggc ggccaattcc gcccctctcc cccccccccc tctccctccc ccccccctaa
     1321 cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc
     1381 caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac
     1441 gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt
     1501 gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg
     1561 caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata
     1621 agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga
     1681 aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt
     1741 accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc
     1801 gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac
     1861 acgatgataa gcttgccaca acccacaagg agacgacctt ccatgaccga gtacaagccc
     1921 acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg
     1981 ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat cgagcgggtc
     2041 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     2101 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     2161 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     2221 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     2281 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     2341 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     2401 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     2461 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gataataggc ggccgctcga
     2521 gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt
     2581 gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct
     2641 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
     2701 ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg
     2761 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     2821 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     2881 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     2941 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
     3001 agtgagaggc cttgacatta taatagattt agcaggaatt gaactaggag tggagcacac
     3061 aggcaaagtt ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     3121 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3181 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3241 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3301 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctgggggcg cgcccctcga
     3361 ggccgccatg gtcatagctg tttgacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3421 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcac gagacaataa
     3481 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     3541 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     3601 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     3661 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     3721 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     3781 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     3841 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     3901 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     3961 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4021 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     4081 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     4141 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     4201 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     4261 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     4321 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     4381 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     4441 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4501 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4561 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4621 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4681 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4741 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4801 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4861 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4921 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4981 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5041 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5101 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5161 ttacggttct ggccttttgc acggaccgcc actgccaatt acctgtggtt tcatttactc
     5221 taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg ttaaatatgt
     5281 actacaaact tagtagttgg aagggctaat tcactcccaa agaagacaag atatccttga
     5341 tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca caccagggcc
     5401 aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga
     5461 taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca
     5521 tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt
     5581 tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat atcgagcttg
     5641 ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg ggactgggga
     5701 gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact gggtctctct
     5761 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     5821 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     5881 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc
     5941 gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct
     6001 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
     6061 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
     6121 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
     6181 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
     6241 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
     6301 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
     6361 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
     6421 taagaccacc gcacagcaac cggccgctga tcttcagacc tggaggagga gatatgaggg
     6481 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     6541 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     6601 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc
     6661 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     6721 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     6781 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     6841 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     6901 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
     6961 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
     7021 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
     7081 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
     7141 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
     7201 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag
     7261 gtggagagag agacagagac agatccattc gattagtgaa cggatctcga cggtatcgcc
     7321 acaaatggca gtattcatcc acaattttaa aagaaagggg gggattgggg ggtacagtgc
     7381 aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca
     7441 aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc cactttggat
     7501 cgataagctt tgcaaagatg gataaagttt taaacagaga ggaatctttg cagctaatgg
     7561 accttctagg tcttgaaagg agtgggaat
//
