LOCUS       pAAV-minBG-CI-mR        4635 bp    DNA     circular SYN 11-SEP-2024
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 4635)
  AUTHORS   Gerard Coughlin
  TITLE     Direct Submission
  JOURNAL   Exported Nov 26, 2024 from SnapGene 8.0.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4635
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     repeat_region   651..791
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4"
     repeat_region   662..791
                     /label=AAV2 ITR
                     /note="color: #a6acb3"
     promoter        complement(794..812)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     misc_feature    813..862
                     /label=stuffer
                     /note="color: #a6acb3"
     promoter        863..881
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     complement(890..914)
                     /label=rBC6-P11
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(917..941)
                     /label=rBC6-P12
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(944..968)
                     /label=rBC6-P13
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(971..995)
                     /label=rBC6-P14
                     /note="color: #a020f0; direction: LEFT"
     misc_feature    1027..1083
                     /label=Minimal B-globin promoter
                     /note="color: #33cccc"
     intron          1168..1300
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
                     /note="color: #------"
     regulatory      1307..1316
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     CDS             1313..2026
                     /codon_start=1
                     /label=mRuby2
                     /note="color: #ff0000"
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     misc_feature    2034..2460
                     /label=W3SL
                     /note="color: #a6acb3"
     misc_feature    2469..2518
                     /label=stuffer
                     /note="color: #a6acb3"
     promoter        2519..2537
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    2538..2557
                     /label=knpI overhang
                     /note="color: #a6acb3"
     repeat_region   2548..2688
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4"
     repeat_region   2548..2677
                     /label=AAV2 ITR
                     /note="color: #a6acb3"
     rep_origin      2763..3218
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        3500..3604
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             join(3605..3673,3674..4465)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 3605 .. 3673 / #ccffcc / signal sequence
                        2: 3674 .. 4465 / #ccffcc
                     Cleavage site after base 3673"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             3605..3673
                     /codon_start=1
                     /label=AmpR
                     /note="color: #993366"
                     /translation="MSIQHFRVALIPFFAAFCLPVFA"
     CDS             3674..4465
                     /codon_start=1
                     /label=AmpR
                     /note="color: #993366"
                     /translation="HPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMS
                     TFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITM
                     SDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMAT
                     TLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIA
                     ALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
      661 gctgcgcgct cgctcgctca ctgaggccgc tcgggcaaag cccgggcgtc gggcgacctt
      721 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
      781 taggggttcc tgccctatag tgagtcgtat tacccggcca ggaaccgtaa aaaggacttg
      841 cccgcgttgc tggcgtttaa cgatttaggt gacactatag aatacacggg tggtaaccgt
      901 attcattgga tttatgtgta tttgccagtg atggcaaatt gccaaggctg ctacggacga
      961 tgtggaagat ggcccaagag catctggaga gacaaggccg cctcgagctt gatatcgaat
     1021 tcctgcagcc cgggctgggc ataaaagtca gggcagagcc atctattgct tacatttgct
     1081 tctagcctgc aggtcgagga gcgcagcctt ccagaagcag agcgcggcgc cttaagctgc
     1141 agaagttggt cgtgaggcac tgggcaggta agtatcaagg ttacaagaca ggtttaagga
     1201 gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct
     1261 attggtctta ctgacatcca ctttgccttt ctctccacag accggtgcca ccatggtgag
     1321 caagggagag gaactcatca aggagaacat gcgcatgaag gtagtcatgg aagggtctgt
     1381 gaatgggcat cagttcaagt gcacagggga aggtgaaggg aatccctaca tgggtactca
     1441 gaccatgagg atcaaggtta ttgaaggtgg ccctctcccg tttgccttcg acattcttgc
     1501 cactagcttc atgtatggca gtcggacctt catcaagtac cccaaaggca ttccagactt
     1561 cttcaaacag tcctttccag agggcttcac atgggaaagg gtgacccgtt acgaggatgg
     1621 tggtgtggtc acagtcatgc aggacactag tctggaggac ggatgtctgg tctatcacgt
     1681 gcaagtgaga ggggtgaact ttccctccaa tgggcctgtg atgcagaaga aaaccaaagg
     1741 gtgggagcca aacacagaga tgatgtatcc tgcagatgga ggactgagag gctatacgca
     1801 catggctctg aaggttgacg gaggaggcca cttgtcatgc tcatttgtga ccacgtacag
     1861 gagcaagaaa actgtcggca acatcaaaat gcctggcata catgcggttg atcaccgctt
     1921 ggaacgactg gaggagtctg acaacgagat gtttgtggta caacgggaac atgcagttgc
     1981 caagtttgct ggccttggcg gaggaatgga tgagctgtac aaatgaacgc gtcataatca
     2041 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     2101 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     2161 tttcattttc tcctccttgt ataaatcctg gttagttctt gccacggcgg aactcatcgc
     2221 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     2281 gtttatttgt gaaatttgtg atgctattgc tttatttgta accatctagc tttatttgtg
     2341 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     2401 acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa
     2461 caaggtaccg ttaaacgcca gcaacgcggg caagtccttt ttacggttcc tggccgggta
     2521 atacgactca ctataggctt gggccgcagg aacccctagt gatggagttg gccactccct
     2581 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     2641 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     2701 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     2761 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2821 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2881 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2941 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     3001 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3061 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     3121 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     3181 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     3241 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     3301 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3361 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     3421 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     3481 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3541 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3601 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3661 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     3721 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     3781 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     3841 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     3901 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     3961 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4021 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4081 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4141 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4201 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4261 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4321 aagccgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4381 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4441 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4501 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4561 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4621 gatcaaagga tcttc
//
