LOCUS       sequence-54135-e        7946 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 7946)
  AUTHORS   Fields RL, Ponzio TA, Kawasaki M, Gainer H
  TITLE     Cell-type specific oxytocin gene expression from AAV delivered 
            promoter deletion constructs into the rat supraoptic nucleus in 
            vivo.
  JOURNAL   PLoS One. 2012;7(2):e32085. Epub 2012 Feb 21.
  PUBMED    22363799
REFERENCE   2  (bases 1 to 7946)
  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: "PLoS One.";
            date: "2012"; volume: "7(2)"; pages: "e32085. Epub 2012 Feb 21"
COMMENT     Sequence Label: pFBOT563
FEATURES             Location/Qualifiers
     source          1..7946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..457
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(89..108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     299..320
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        484..588
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(589..657,658..1449)
                     /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(807..826)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1620..2208
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2109..2128
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  2511..2735
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2802..3335)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3524..3529,3530..3546,3547..3552))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     repeat_region   3917..4057
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3928..4057
                     /label=AAV2 ITR
     regulatory      5477..5486
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5483..5485,5486..5488,5489..6202)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5483..5485,5486..5488,5489..6199)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5528..5549)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5789..5808)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6136..6157
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     repeat_region   7155..7295
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7155..7284
                     /label=AAV2 ITR
     polyA_signal    7437..7571
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7474..7493)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7528..7547
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  7600..7765
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
       61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
      301 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
      361 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
      421 aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt tcggggaaat
      481 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
      541 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
      601 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
      661 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
      721 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
      781 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
      841 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
      901 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
      961 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     1021 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     1081 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     1141 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     1201 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     1261 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     1321 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     1381 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     1441 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     1501 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     1561 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1621 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1681 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     1741 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     1801 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     1861 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     1921 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     1981 tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg
     2041 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2101 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2161 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2221 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     2281 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     2341 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     2401 cggtattttc tccttacgca tctgtgcggt atttcacacc gcagaccagc cgcgtaacct
     2461 ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg tgtgggcgga
     2521 caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag
     2581 acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt
     2641 tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga
     2701 ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa tttaccgaac
     2761 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg
     2821 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
     2881 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
     2941 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact
     3001 gcgagatcat agatatagat ctcactacgc ggctgctcaa acctgggcag aacgtaagcc
     3061 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta
     3121 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct
     3181 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
     3241 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
     3301 ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata acatcaaaca
     3361 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtacaaaaaa
     3421 acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa
     3481 ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac gaaccgaaca
     3541 ggcttatgtc aactgggttc gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac
     3601 cttgggcagc agcgaagtcg aggcatttct gtcctggctg gcgaacgagc gcaaggtttc
     3661 ggtctccacg catcgtcagg cattggcggc cttgctgttc ttctacggca aggtgctgtg
     3721 cacggatctg ccctggcttc aggagatcgg aagacctcgg ccgtcgcggc gcttgccggt
     3781 ggtgctgacc ccggatgaag tggttcgcat cctcggtttt ctggaaggcg agcatcgttt
     3841 gttcgcccag gactctagct atagttctag tggttggcta cattattgaa gcatttatca
     3901 gggttattgt ctcagacctg caggcagctg cgcgctcgct cgctcactga ggccgcccgg
     3961 gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc
     4021 agagagggag tggccaactc catcactagg ggttccttgt agttaatgat taacccgcca
     4081 tgctacttat ctacgtagcc atgctctaga gctcaagctt atcgataccg tctacacagc
     4141 aggttctaat acagagtttt aaatactagt gtaattttcc ttttgctgta atttttcttt
     4201 ctttttattt taatttggtc tgttaactct gttttggttt tgatctctac tgagactttg
     4261 ttttacgggc tagttagaag agctgaggtg cattcagaga ttgaacaaga cgccatcttt
     4321 ttccccatct aagtttcagg tccatgtaag ggctccctca ctcactggcc ctatcctgcc
     4381 ttattctgag atattggata tctgtgaaaa acagctcctg gctagggcgc acctccaacc
     4441 cctcccaagt ctctctagcc tcttgtagcc taggccaccc cttccaggct gcttctcttt
     4501 tgagttccag gtcattagca gagacgatga ccttgaccct agcccagacc ctgcaaatga
     4561 agggcctgcc tctaaacagc gtggaacaat ttcacccaag agaccttctg tgaccagtca
     4621 tgctgtcacc ctctttagac agtgctccac catggcagtg ccagacataa aaaggtcggt
     4681 ctgggccgga gaaaccatca cctacagcgg atctcagact gagcaccatc gccatggcct
     4741 gccccagtct cgcttgctgc ctgcttggct tactggctct gacctcggcc tgctacatcc
     4801 agaactgccc cctgggcggc aagagggctg tgctggacct ggatatgcgc aaggttagtc
     4861 tccccgaccc tgtcccttcc cttcccgttc tggcgatgct aaggaccaga gaagctctcc
     4921 cacctacaga gagcattccc gcacacttgc cagccctacc aaggcctcgc gtgggaaccc
     4981 agggctttgg gaagtgttag gctccctctt gacgccgtga aggtaacgac aatgccggag
     5041 cacccactgc ccctcgctct gccacagtcc ggattcggat tgtgcacggc gcccacccgc
     5101 atccttcccc acagtgtctc ccctgcggcc cgggcggcaa aggacgctgc ttcggaccaa
     5161 gcatctgctg cgcggacgag ctgggctgct tcgtgggcac cgccgaggcg ctgcgctgcc
     5221 aggaggagaa ctacctgcct tcgccctgcc agtctggcca gaagccctgc gggagcggag
     5281 gccgctgcgc cgccacaggc atctgctgca gcccgggtga gcaggagggg gcccagcagg
     5341 tgacccggca aggagccgtc gggtttgcag ctcagaacac tgacccattt ctcttgcaga
     5401 tggctgccgc acagaccccg cctgcgaccc tgagtctgcc ttctcggagt tgcggcccgg
     5461 gatccaccgg ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
     5521 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     5581 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     5641 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     5701 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     5761 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     5821 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     5881 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     5941 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     6001 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     6061 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     6121 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     6181 catggacgag ctgtacaagt aaagcggccg caagctgagc ccactttctg ggaatacctt
     6241 tagcgcgctt ccttcgttcc ccatggccac tgccagaaaa aaaaaaaaaa aagaaaagaa
     6301 aagaaaagaa aagaaaaata aagtagattt cctcttcaaa cttgactggt gtctaattgt
     6361 cggaaacggg agggaggaaa ggcaccggga acgccgtgct cttggcatct tgtagcaaaa
     6421 acagtccagc aggctgctcg gggaccgagg ggtgctctat gggctgccat ctcccattag
     6481 ccccatttcc atttcaactg ggctgggcct gatagactta agagaagaat ttggaagtcc
     6541 aaacagcaac ggtggacttc tgtgggaaag ggcaacgcct aacaggggac tgaggagcct
     6601 aacaaggcag gctaggaggc aggggaacta gttcagtcac ttccatatct gagagggggc
     6661 tttagctgtg acctcagagg ttgagggggc aatcctggcc tagaaggcag ggtcttggcc
     6721 aggggagggt agttacagaa ccataaggtt tccacagagc tgaattctca cgcaggcccc
     6781 aaggaaacca cagctggaaa gatcaagaac caactggcct ctcctctgct gtccccaggg
     6841 tcgggcctga gctgggggtg ggggtggggg gcgggggggg ggctgcccgc ctctgcagga
     6901 cagcgtgctc tggggagaca tgggatctaa gagccagtgg gcacaagtga ccacaaagca
     6961 cggagcccac agacaagcag aaatccacaa tctatactgc tcccttactg ctggattaaa
     7021 ataccgccct tgccacgaag cgcgcctcac ttggctgcca gtgttctcag cagcagtaat
     7081 gggcagaagt gagggcatgg ctctagagca tggctacgta gataagtagc atggcgggtt
     7141 aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     7201 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     7261 ctcagtgagc gagcgagcgc gcagctgcct gcaggcatgc aagctgtaag cttgtcgaga
     7321 agtactagag gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     7381 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     7441 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     7501 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     7561 atgtctggat ctgatcactg atatcgccta ggagatccga accagataag tgaaatctag
     7621 ttccaaacta ttttgtcatt tttaattttc gtattagctt acgacgctac acccagttcc
     7681 catctatttt gtcactcttc cctaaataat ccttaaaaac tccatttcca cccctcccag
     7741 ttcccaacta ttttgtccgc ccacagcggg gcatttttct tcctgttatg tttttaatca
     7801 aacatcctgc caactccatg tgacaaaccg tcatcttcgg ctactttttc tctgtcacag
     7861 aatgaaaatt tttctgtcat ctcttcgtta ttaatgtttg taattgactg aatatcaacg
     7921 cttatttgca gcctgaatgg cgaatg
//
