LOCUS       40924_40192        5201 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSI-AGL10, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5201)
  AUTHORS   Mansouri M, Bellon-Echeverria I, Rizk A, Ehsaei Z, Cianciolo 
            Cosentino C, Silva CS, Xie Y, Boyce FM, Davis MW, Neuhauss SC, 
            Taylor V, Ballmer-Hofer K, Berger I, Berger P.
  TITLE     Highly efficient baculovirus-mediated multigene delivery in primary 
            cells
  JOURNAL   Nat Commun 7, 11529 (2016)
  PUBMED    27143231
REFERENCE   2  (bases 1 to 5201)
  AUTHORS   Mansouri M, Berger I, Berger P.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-MAR-2016) Laboratory of Biomolecular Research, Paul 
            Scherrer Institute, OFLG-110, Villigen psi 5232, Switzerland
REFERENCE   3  (bases 1 to 5201)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5201)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Commun 
            7, 11529 (2016)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-MAR-2016) Laboratory of Biomolecular Research, Paul Scherrer 
            Institute, OFLG-110, Villigen psi 5232, Switzerland"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Ape v. v. v2.0.49
            Sequencing Technology :: 454; Illumina
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5201
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      190..778
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     mobile_element  complement(913..1136)
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1206..1736)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(1925..1953)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(2024..2057)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        2149..2452
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2453..2656
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        2701..2719
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2781..3497
                     /codon_start=1
                     /label=mEYFP
                     /note="enhanced YFP with monomerizing A206K mutation
                     (Zacharias et al., 2002)"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     protein_bind    3556..3577
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3592..3622
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3630..3646
                     /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     3654..3670
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             3697..3719
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3844..4551
                     /codon_start=1
                     /label=GFP
                     /note="Aequorea victoria green fluorescent protein"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLCYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELY"
     polyA_signal    4744..4968
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     mobile_element  complement(5036..5201)
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 accggttgac ttgggtcaac tgtcagacca agtttactca tatatacttt agattgattt
       61 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
      121 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
      181 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
      241 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
      301 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
      361 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
      421 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
      481 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
      541 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
      601 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
      661 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
      721 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
      781 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
      841 ctttcctgcg ttatcccctg attgacttgg gtcactcttc ctgtggatgc gcagatgccc
      901 tgcgtaagcg ggtgtgggcg gacaataaag tcttaaactg aacaaaatag atctaaacta
      961 tgacaataaa gtcttaaact agacagaata gttgtaaact gaaatcagtc cagttatgct
     1021 gtgaaaaagc atactggact tttgttatgg ctaaagcaaa ctcttcattt tctgaagtgc
     1081 aaattgcccg tcgtattaaa gaggggcgtg gccaagggca tgtaaagact atattcgcgg
     1141 cgttgtgaca atttaccgaa caactccgcg gccgggaagc cgatctcggc ttgaacgaat
     1201 tgttaggtgg cggtacttgg gtcgatatca aagtgcatca cttcttcccg tatgcccaac
     1261 tttgtataga gagccactgc gggatcgtca ccgtaatctg cttgcacgta gatcacataa
     1321 gcaccaagcg cgttggcctc atgcttgagg agattgatga gcgcggtggc aatgccctgc
     1381 ctccggtgct cgccggagac tgcgagatca tagatataga tctcactacg cggctgctca
     1441 aacttgggca gaacgtaagc cgcgagagcg ccaacaaccg cttcttggtc gaaggcagca
     1501 agcgcgatga atgtcttact acggagcaag ttcccgaggt aatcggagtc cggctgatgt
     1561 tgggagtagg tggctacgtc tccgaactca cgaccgaaaa gatcaagagc agcccgcatg
     1621 gatttgactt ggtcagggcc gagcctacat gtgcgaatga tgcccatact tgagccacct
     1681 aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctgcg taacatcgtt
     1741 gctgctccat aacatcaaac atcgacccac ggcgtaacgc gcttgctgct tggatgcccg
     1801 aggcatagac tgtacaaaaa aacagtcata acaagccatg aaaaccgcca ctgcgccgtt
     1861 accaccgctg cgttcggtca aggttctgga ccagttgcgt gagcgcatac gctacttgca
     1921 ttacagttta cgaaccgaac aggcttatgt caactgggtt cgtgccttca tccgtttcca
     1981 cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc gccataactt cgtatagcat
     2041 acattatacg aagttatctg taactataac ggtcctaagt ggcgcgccta gttattaata
     2101 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     2161 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     2221 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     2281 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     2341 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     2401 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     2461 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     2521 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     2581 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     2641 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat
     2701 taatacgact cactataggg agacccaagc tggctagcgt ttaaacttaa gcttggtacc
     2761 gagctcaaag gagatatacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     2821 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     2881 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     2941 tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta cggcctgatg tgcttcgccc
     3001 gctaccccga ccacatgaag cgtcacgact tcttcaagtc cgccatgccc gaaggctacg
     3061 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     3121 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     3181 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     3241 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     3301 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     3361 tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa gaccccaacg
     3421 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actcacggca
     3481 tggacgagct gtacaaggga tcaggatcag gatcaatggg aagagctgga aagcgggcag
     3541 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     3601 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     3661 cagctatgac catgattacg aatttctaga aataattttg tttaacttta agaaggagat
     3721 atacatatgg ctagaatcca cagcagatac aaagaacttc ttgccaaaag agcagagctt
     3781 caaagaaaag tagaggaact tcagagagaa atttccaacc gctcaacctc ctcttcagag
     3841 tctagcaaag gagaagaact cttcactgga gttgtcccaa ttcttgttga attagatggt
     3901 gatgttaacg gccacaagtt ctctgtcagt ggagagggtg aaggtgatgc aacatacgga
     3961 aaacttaccc tgaagttcat ctgcactact ggcaaactgc ctgttccatg gccaacacta
     4021 gtcactactc tgtgctatgg tgttcaatgc ttttcaagat acccggatca tatgaaacgg
     4081 catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaaggac catcttcttc
     4141 aaagatgacg gcaactacaa gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt
     4201 aatagaatcg agttaaaagg tattgacttc aaggaagatg gcaacattct gggacacaaa
     4261 ttggaataca actataactc acacaatgta tacatcatgg cagacaaaca aaagaatgga
     4321 atcaaagtga acttcaagac ccgccacaac attgaagatg gaagcgttca actagcagac
     4381 cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac
     4441 ctgtccacac aatctgccct ttcgaaagat cccaacgaaa agagagacca catggtcctt
     4501 cttgagtttg taacagctgc tgggattaca catggcatgg atgaactgta caactgagta
     4561 aattacattt tatttacaat cactcgacga agacttgatc acccgggatc tcgagccatg
     4621 gtgctagcag ctgatgcata gcatgcggta ccgggagatg ggggaggcta actgaaacac
     4681 ggaagggctc ttcctgaccg ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct
     4741 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     4801 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     4861 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     4921 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     4981 aaagaaccag ctggggctct aggttaatta acctagggta tacccatcta attggaacca
     5041 gataagtgaa atctagttcc aaactatttt gtcattttta attttcgtat tagcttacga
     5101 cgctacaccc agttcccatc tattttgtca ctcttcccta aataatcctt aaaaactcca
     5161 tttccacccc tcccagttcc caactatttt gtccgcccac a
//
