LOCUS       40924_38853        4369 bp DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4369)
  AUTHORS   Gray JT, Zolotukhin S
  TITLE     Design and Construction of Functional AAV Vectors.
  JOURNAL   Methods Mol Biol. 2011;807:25-46.
  PUBMED    22034025
REFERENCE   2  (bases 1 to 4369)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4369)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Methods Mol
            Biol."; date: "2011"; volume: "807"; pages: "25-46"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        571..774
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     primer_bind     771..795
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          823..888
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             942..1658
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     1705..1724
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1974..1990)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2199..2218)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     primer_bind     2318..2340
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2378..2396)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     promoter        2464..2568
                     /label=AmpR promoter
     CDS             2569..3426
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3600..4188
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4342..4359
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct
      241 cgagatctag aaagcttccc ggggggatct gggccactcc ctctctgcgc gctcgctcgc
      301 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
      361 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctggaggg
      421 gtggagtcgt gacctaggca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      481 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      541 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      601 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      661 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      721 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
      781 gtgaaccgtc agatcgcctg gagacgccat ccggactcta aggtaaatat aaaattttta
      841 agtgtataat gtgttaaact actgattcta attgtttctc tcttttagat tccaaccttt
      901 ggaactgaat tccgcgggcc cgggatccac cggtcgccac catggtgagc aagggcgagg
      961 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1021 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     1081 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     1141 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     1201 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     1261 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1321 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     1381 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     1441 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     1501 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     1561 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1621 ccgccgggat cactctcggc atggacgagc tgtacaagta aagcggccgc taggcctcac
     1681 ctgcgatctc gatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1741 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     1801 ggggaggtgt gggaggtttt ttaaactagt ccactccctc tctgcgcgct cgctcgctca
     1861 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     1921 gcgagcgagc gcgcagagag ggacagatcc gggcccgcat gcgtcgacaa ttcactggcc
     1981 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     2041 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     2101 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
     2161 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
     2221 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     2281 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     2341 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     2401 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     2461 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2521 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2581 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2641 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2701 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2761 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     2821 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2881 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2941 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     3001 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     3061 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     3121 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     3181 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3241 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3301 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3361 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3421 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     3481 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     3541 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3601 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3661 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3721 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     3781 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3841 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3901 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3961 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4021 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4081 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4141 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4201 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4261 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4321 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaag
//
