LOCUS       Exported                5980 bp DNA     circular SYN 04-SEP-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pAAV hSYN hPOMC1-26-sbGluc-p2a-Dtomato
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5980)
  AUTHORS   Hochgeschwender
  TITLE     Direct Submission
  JOURNAL   Exported Sep 14, 2021 from SnapGene 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5980
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   5..145
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     promoter        190..637
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             662..739
                     /codon_start=1
                     /label=hPOMC1-26
                     /translation="MPRSCCSRSGALLLALLLQASMEVRG"
     CDS             740..1243
                     /codon_start=1
                     /label=sbGluc w/o atg
                     /translation="KPTENNEDFNIVAVASNFATTDLDADRGKLPGKKLPLEVLKELEA
                     NARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESAQGGIGEAIVDIPEIPGF
                     KDLEPLEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQRCATFASKIQGQVDKIK
                     GAGGD"
     CDS             740..1003
                     /codon_start=1
                     /label=N-sbGluc w/o signal sequence
                     /translation="KPTENNEDFNIVAVASNFATTDLDADRGKLPGKKLPLEVLKELEA
                     NARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESAQ"
     CDS             1004..1243
                     /codon_start=1
                     /label=C-sbGluc
                     /translation="GGIGEAIVDIPEIPGFKDLEPLEQFIAQVDLCVDCTTGCLKGLAN
                     VQCSDLLKKWLPQRCATFASKIQGQVDKIKGAGGD"
     CDS             1250..1315
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="GSGATNFSLLKQAGDVEENPGP"
     CDS             1334..2035
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     CDS             2063..2080
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2111..2699
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(2582..2593)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2731..3207
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     repeat_region   3247..3387
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      3462..3917
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4199..4303
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4304..5164
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5335..5923
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg
       61 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
      121 gccaactcca tcactagggg ttcctgcggc cgcacgcgtg tgtctagact gcagagggcc
      181 ctgcgtatga gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct
      241 gacgaccgac cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc
      301 cctatcagag agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc
      361 agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac
      421 tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg
      481 tcgcgtccgc gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg
      541 gcacgggcgc gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt
      601 gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcg agaaggtacc ggatccccac
      661 catgccgaga tcgtgctgca gccgctcggg ggccctgttg ctggccttgc tgcttcaggc
      721 ctccatggaa gtgcgtggca agcccaccga gaacaacgaa gacttcaaca tcgtggccgt
      781 ggccagcaac ttcgcgacca cggatctcga tgctgaccgc gggaagttgc ccggcaagaa
      841 gctgccgctg gaggtgctca aagagctgga agccaatgcc cggaaagctg gctgcaccag
      901 gggctgtctg atctgcctgt cccacatcaa gtgcacgccc aagatgaaga agttcatccc
      961 aggacgctgc cacacctacg aaggcgacaa agagtccgca cagggcggca taggcgaggc
     1021 gatcgtcgac attcctgaga ttcctgggtt caaggacttg gagcccctgg agcagttcat
     1081 cgcacaggtc gatctgtgtg tggactgcac aactggctgc ctcaaagggc ttgccaacgt
     1141 gcagtgttct gacctgctca agaagtggct gccgcaacgc tgtgcgacct ttgccagcaa
     1201 gatccagggc caggtggaca agatcaaggg ggccggtggt gacctcgagg gaagcggagc
     1261 cactaacttc tccctgttga aacaagcagg ggatgtcgaa gagaatcccg ggccagcggc
     1321 cgcacccggg ccaatggtga gcaagggcga ggaggtcatc aaagagttca tgcgcttcaa
     1381 ggtgcgcatg gagggctcca tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     1441 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc
     1501 cttcgcctgg gacatcctgt ccccccagtt catgtacggc tccaaggcgt acgtgaagca
     1561 ccccgccgac atccccgatt acaagaagct gtccttcccc gagggcttca agtgggagcg
     1621 cgtgatgaac ttcgaggacg gcggtctggt gaccgtgacc caggactcct ccctgcagga
     1681 cggcacgctg atctacaagg tgaagatgcg cggcaccaac ttcccccccg acggccccgt
     1741 aatgcagaag aagaccatgg gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg
     1801 cgtgctgaag ggcgagatcc accaggccct gaagctgaag gacggcggcc actacctggt
     1861 ggagttcaag accatctaca tggccaagaa gcccgtgcaa ctgcccggct actactacgt
     1921 ggacaccaag ctggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     1981 gcgctccgag ggccgccacc acctgttcct gtacggcatg gacgagctgt acaagtaaca
     2041 tatgctctag acaattgccg gtcatcatca ccatcaccat tgagtttaat tcgatatcaa
     2101 gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2161 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2221 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     2281 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     2341 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     2401 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     2461 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     2521 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     2581 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     2641 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     2701 tcgataccga gcgctgctcg agagatctac gggtggcatc cctgtgaccc ctccccagtg
     2761 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta
     2821 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg
     2881 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa
     2941 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa
     3001 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc
     3061 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc
     3121 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg
     3181 tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc
     3241 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     3301 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     3361 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     3421 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     3481 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3541 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3601 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3661 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3721 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3781 aacaacactc aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc
     3841 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     3901 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3961 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4021 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4081 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     4141 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4201 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4261 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4321 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4381 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4441 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4501 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4561 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4621 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4681 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4741 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4801 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4861 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4921 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4981 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5041 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5101 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5161 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5221 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5281 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5341 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5401 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5461 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     5521 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5581 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5641 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5701 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5761 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5821 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5881 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5941 ctttttacgg ttcctggcct tttgctggcc ttttgctcac
//
