LOCUS       40924_40197        5192 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSI-AGR10, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5192)
  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 5192)
  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 5192)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5192)
  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..5192
                     /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..3488
                     /codon_start=1
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     protein_bind    3547..3568
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3583..3613
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3621..3637
                     /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     3645..3661
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             3688..3710
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3835..4542
                     /codon_start=1
                     /label=GFP
                     /note="Aequorea victoria green fluorescent protein"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLCYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELY"
     polyA_signal    4735..4959
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     mobile_element  complement(5027..5192)
                     /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 ggataacatg gccatcatca
     2821 aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga
     2881 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
     2941 ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct
     3001 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg
     3061 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc
     3121 aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact
     3181 tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc
     3241 ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg
     3301 acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc
     3361 tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca
     3421 ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc
     3481 tgtacaaggc tcaaaggaga tataccatgg gaagagctgg aaagcgggca gtgagcgcaa
     3541 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     3601 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     3661 ccatgattac gaatttctag aaataatttt gtttaacttt aagaaggaga tatacatatg
     3721 gctagaatcc acagcagata caaagaactt cttgccaaaa gagcagagct tcaaagaaaa
     3781 gtagaggaac ttcagagaga aatttccaac cgctcaacct cctcttcaga gtctagcaaa
     3841 ggagaagaac tcttcactgg agttgtccca attcttgttg aattagatgg tgatgttaac
     3901 ggccacaagt tctctgtcag tggagagggt gaaggtgatg caacatacgg aaaacttacc
     3961 ctgaagttca tctgcactac tggcaaactg cctgttccat ggccaacact agtcactact
     4021 ctgtgctatg gtgttcaatg cttttcaaga tacccggatc atatgaaacg gcatgacttt
     4081 ttcaagagtg ccatgcccga aggttatgta caggaaagga ccatcttctt caaagatgac
     4141 ggcaactaca agacacgtgc tgaagtcaag tttgaaggtg atacccttgt taatagaatc
     4201 gagttaaaag gtattgactt caaggaagat ggcaacattc tgggacacaa attggaatac
     4261 aactataact cacacaatgt atacatcatg gcagacaaac aaaagaatgg aatcaaagtg
     4321 aacttcaaga cccgccacaa cattgaagat ggaagcgttc aactagcaga ccattatcaa
     4381 caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta cctgtccaca
     4441 caatctgccc tttcgaaaga tcccaacgaa aagagagacc acatggtcct tcttgagttt
     4501 gtaacagctg ctgggattac acatggcatg gatgaactgt acaactgagt aaattacatt
     4561 ttatttacaa tcactcgacg aagacttgat cacccgggat ctcgagccat ggtgctagca
     4621 gctgatgcat agcatgcggt accgggagat gggggaggct aactgaaaca cggaagggct
     4681 cttcctgacc gctcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc
     4741 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     4801 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     4861 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     4921 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     4981 gctggggctc taggttaatt aacctagggt atacccatct aattggaacc agataagtga
     5041 aatctagttc caaactattt tgtcattttt aattttcgta ttagcttacg acgctacacc
     5101 cagttcccat ctattttgtc actcttccct aaataatcct taaaaactcc atttccaccc
     5161 ctcccagttc ccaactattt tgtccgccca ca
//
