LOCUS       sequence-47850-e        7794 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7794)
  AUTHORS   Watabe-Uchida M, Zhu L, Ogawa SK, Vamanrao A, Uchida N
  TITLE     Whole-brain mapping of direct inputs to midbrain dopamine neurons.
  JOURNAL   Neuron. 2012 Jun 7;74(5):858-73.
  PUBMED    22681690
REFERENCE   2  (bases 1 to 7794)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Neuron."; 
            date: "2012-06-7"; volume: "74(5)"; pages: "858-73"
COMMENT     Sequence Label: pAAV-CA-FLEX-RG
FEATURES             Location/Qualifiers
     source          1..7794
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     enhancer        235..538
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        541..816
                     /label=chicken beta-actin promoter
     intron          817..1825
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1749..1771
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     misc_feature    3921..4509
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3974..3994)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4392..4403)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4541..5017
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4740..4759)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5057..5197
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5057..5186
                     /label=AAV2 ITR
     rep_origin      5272..5727
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5359..5378)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5569..5590
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5744..5763)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5863..5885
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5923..5941)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6009..6113
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6114..6182,6183..6974)
                     /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"
     primer_bind     complement(6332..6351)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7145..7733
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7634..7653
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagct gggctgcagg aatctagtta
      181 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      241 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      301 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      361 ggactattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      421 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      481 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggg
      541 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa
      601 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg
      661 cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg
      721 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
      781 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgtt gccttcgccc
      841 cgtgccccgc tccgcgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
      901 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa
      961 tgacggctcg tttcttttct gtggctgcgt gaaagcctta aagggctccg ggagggccct
     1021 ttgtgcgggg gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc
     1081 gtgcggcccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg
     1141 ctccgcgtgt gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc gggggggctg
     1201 cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag ggggtgtggg
     1261 cgcggcggtc gggctgtaac ccccccctgc acccccctcc ccgagttgct gagcacggcc
     1321 cggcttcggg tgcggggctc cgtacggggc gtggcgcggg gctcgccgtg ccgggcgggg
     1381 ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc tcgggccggg gagggctcgg
     1441 gggaggggcg cggcggcccc ggagcgccgg cggctgtcga ggcgcggcga gccgcagcca
     1501 ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc aaatctggcg
     1561 gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcgggcg aagcggtgcg
     1621 gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc cgcggtcccc
     1681 ttctccatct ccagcctcgg ggctgccgca gggggacggc tgccttcggg ggggacgggg
     1741 cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt
     1801 tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttgttg tgctgtctca
     1861 tcattttggc aaagaattga tttcgatacg tacggttgaa ttcattacct cgtataggat
     1921 actttatacg aagttatgca gaatggtagc tggattgtaa gtactattag caatatgaaa
     1981 cctcttaata acctcgtata gcatacatta tacgaagtaa tagatctaag cttgattaat
     2041 gaaaatctca tgactcttga atctacccag ttgcatggag ttcaaggagg actattgaac
     2101 ccaggttttt caaaaagaac cccccaaggg gaggtgatct tcaggacttg gatcgttgaa
     2161 aggacggcca gtccttacag tctggtctca cccccactct tgtgtgattc ccatgaagat
     2221 atgatcttcc cgctttgggg agtgactgac acctccctcc ctgtccctct gagattgtgt
     2281 tgcgtaggtt ctgatcgatt gactcttcta caacatgtca tcaggaaaat tatcaacatc
     2341 aaggcagtca gggcccctgc actcagtaat acatacttcc cccagttcgg gagacccaag
     2401 tcaactcctg agacctgatt gtgcacatcg ggaaggtgaa cttcaacaaa atcctcagcc
     2461 tcgtcaccgt ccttgaaaac ggtagacggg tctgccaggg ggtgcacaag ggggataacc
     2521 gaggattcca acaactccat atgttgctgg aggagggatg attgcatctc tgggattaag
     2581 acattgccgt caggtcctaa tattatacca ttgaaaaaca ccccgttcac atgaggatga
     2641 cacctccccc caactcttaa acaccctttt gaagggagga tctcattcca agttctgact
     2701 gacttgtagt gagcatcggc ttccatcaag gtcttgttga atatggtata tgcttttcca
     2761 aacccaggga caagttttct taaatgactg agacgtctga aactcactga cttggttgtc
     2821 atgatggact ctagtgcatc cagacactcc tctctcttcc tgaccaactc ctctacaaca
     2881 aggtgctcaa tttcgtctga gcgaaagtcg tgcaggttca ccaacttatc gggagggcac
     2941 catttggttt catttgatgt ttgcatcgag acccatgttc catccataag tctaagtcct
     3001 agaactccac ataacttgag tttgcatgct ccttttaaag acttatatag gcctctttca
     3061 tctacaaagc cgcaagtctc actccctttg gatgctctct tccctctact attggtaaaa
     3121 atgtcacaag acatccctag tctcggattc tcgggcatcc aaatggtgta atcgtggtta
     3181 gtggagcagt aggtagaaga caccgctact cctgagcact tcccgctagg gaagaccctc
     3241 gagtgaaggg atctgtcata tgggtccaaa tctgccacac ttggagatat gataacgaga
     3301 gactccttgg tggtttttac agttcgaagc cagcggtagt cagggtacgg attgtgtaga
     3361 gactcttcat atctggggtc accggccatc ttccagttgt acgcggctct acatgcatct
     3421 ggtgttgggc ggaaatgctt tcttttgaac gtggttgtga cataaccaac gaagttagtg
     3481 taggtttcag cctccgtcac aacgcctgtg caagtgaacc cgttcacttt tatggctaag
     3541 atgtatccaa ctttaagttc catgtaggag aaccctgaca ggttggtgca tccttcgtcc
     3601 tccactacca aattgtttgg gcagctgagg tgatgtatgt caatcggact ccagggacca
     3661 agcttgtctg gtatcgtgta aatagggaat ttcccaaaac acaatggaaa aaccagaagg
     3721 ggtacaaaca ggagagcctg aggaaccatc tttccctcgt gccgaattcg atatcgtcga
     3781 cataacttcg tataaagtat cctatacgag gtaatttgcc ttaacccaga aattatcagt
     3841 actattcttt agaatggtgc aaagaattac ttcgtataat gtatgctata cgaggttatc
     3901 gcatgctaga gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg
     3961 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     4021 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     4081 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     4141 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     4201 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     4261 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     4321 cgtcctttcc ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct
     4381 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     4441 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     4501 cctccccgca tcgataccga gcgctgctcg agagatctac gggtggcatc cctgtgaccc
     4561 ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct
     4621 aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg
     4681 gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt
     4741 ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc
     4801 ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat
     4861 gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag
     4921 gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg
     4981 attacaggcg tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg
     5041 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     5101 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     5161 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     5221 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     5281 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     5341 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     5401 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     5461 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     5521 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     5581 tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta taagggattt
     5641 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     5701 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     5761 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     5821 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     5881 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     5941 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     6001 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     6061 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     6121 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     6181 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     6241 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     6301 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     6361 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     6421 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     6481 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     6541 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     6601 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     6661 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     6721 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     6781 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     6841 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     6901 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6961 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     7021 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     7081 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     7141 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     7201 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     7261 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     7321 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     7381 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     7441 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     7501 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     7561 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     7621 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     7681 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     7741 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
