LOCUS       MB35paav-cag-egf        5464 bp    DNA     circular SYN 28-JAN-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5464)
  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..5464
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     enhancer        1008..1387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     regulatory      2128..2137
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     CDS             join(2134..2136,2137..2139,2140..2142,2143..2856)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 4 segments:
                        1: 2134 .. 2136 / #05fd14
                        2: 2137 .. 2139 / #05fd14
                        3: 2140 .. 2142 / #05fd14 / 1a
                        4: 2143 .. 2856 / #05fd14"
                     /translation="MAVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLT
                     LKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKD
                     DGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGI
                     KVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVL
                     LEFVTAAGITLGMDELYK"
     misc_feature    2863..3289
                     /label=W3SL
                     /note="color: #a6acb3"
     misc_feature    3298..3347
                     /label=stuffer
                     /note="color: #a6acb3"
     promoter        3348..3366
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    3367..3386
                     /label=knpI overhang
                     /note="color: #a6acb3"
     repeat_region   3377..3517
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4"
     repeat_region   3377..3506
                     /label=AAV2 ITR
                     /note="color: #a6acb3"
     rep_origin      3592..4047
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        4329..4433
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             join(4434..4502,4503..5294)
                     /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: 4434 .. 4502 / #ccffcc / signal sequence
                        2: 4503 .. 5294 / #ccffcc
                     Cleavage site after base 4502"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             4434..4502
                     /codon_start=1
                     /label=AmpR
                     /note="color: #993366"
                     /translation="MSIQHFRVALIPFFAAFCLPVFA"
     CDS             4503..5294
                     /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 cctcgaggac attgattatt
     1021 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     1081 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     1141 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     1201 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     1261 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     1321 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     1381 taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc
     1441 acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg
     1501 gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg gggcggggcg aggcggagag
     1561 gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc
     1621 ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc
     1681 ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg
     1741 cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct
     1801 tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag gggctccggg
     1861 agcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc
     1921 cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg
     1981 ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat
     2041 gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct
     2101 catcattttg gcaaagaatt gaccggtgcc accatggccg tgagcaaggg cgaggagctg
     2161 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     2221 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     2281 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     2341 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     2401 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     2461 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     2521 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     2581 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     2641 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     2701 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     2761 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     2821 gggatcactc tcggcatgga cgagctgtac aagtaggaat tcataatcaa cctctggatt
     2881 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     2941 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     3001 cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc gcctgccttg
     3061 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg tttatttgtg
     3121 aaatttgtga tgctattgct ttatttgtaa ccatctagct ttatttgtga aatttgtgat
     3181 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     3241 attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaac aaggtaccgt
     3301 taaacgccag caacgcgggc aagtcctttt tacggttcct ggccgggtaa tacgactcac
     3361 tataggcttg ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     3421 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     3481 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     3541 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     3601 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3661 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3721 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3781 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     3841 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3901 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     3961 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4021 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     4081 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     4141 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     4201 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     4261 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     4321 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4381 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4441 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4501 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4561 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4621 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4681 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4741 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4801 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4861 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4921 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4981 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5041 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5101 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtgga agccgcggta
     5161 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5221 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5281 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5341 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5401 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5461 cttc
//
