LOCUS       sequence-54133-e        8799 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 8799)
  AUTHORS   Fields RL, House SB, Gainer H
  TITLE     Regulatory domains in the intergenic region of the oxytocin and 
            vasopressin genes that control their hypothalamus-specific 
            expression in vitro.
  JOURNAL   J Neurosci. 2003 Aug 27;23(21):7801-9.
  PUBMED    12944509
REFERENCE   2  (bases 1 to 8799)
  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: "J 
            Neurosci."; date: "2003-08-27"; volume: "23(21)"; pages: "7801-9"
COMMENT     Sequence Label: pOTIII.EFGP.IGR3.6
FEATURES             Location/Qualifiers
     source          1..8799
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(129..197,198..989)
                     /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(347..366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1649..1668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1902..1919
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2072..2077,2078..2095,2096..2102)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2115..2137
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2134..2150
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2169..2189
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2171..2189
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2230..2246
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2230..2246
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     regulatory      3634..3643
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3640..3642,3643..3645,3646..4359)
                     /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(3640..3642,3643..3645,3646..4356)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3685..3706)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3946..3965)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4293..4314
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(8155..8174)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8156..8174)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8184..8201)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8184..8200)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8193..8215)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8342..8797
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8429..8448)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8639..8660
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     2161 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctccac cgcggtggcg
     2221 gccggccgct ctagaactag tggatccccc gggctgcagg aattcgatat caagcttatc
     2281 gataccgtct acacagcagg ttctaataca gagttttaaa tactagtgta attttccttt
     2341 tgctgtaatt tttctttctt tttattttaa tttggtctgt taactctgtt ttggttttga
     2401 tctctactga gactttgttt tacgggctag ttagaagagc tgaggtgcat tcagagattg
     2461 aacaagacgc catctttttc cccatctaag tttcaggtcc atgtaagggc tccctcactc
     2521 actggcccta tcctgcctta ttctgagata ttggatatct gtgaaaaaca gctcctggct
     2581 agggcgcacc tccaacccct cccaagtctc tctagcctct tgtagcctag gccacccctt
     2641 ccaggctgct tctcttttga gttccaggtc attagcagag acgatgacct tgaccctagc
     2701 ccagaccctg caaatgaagg gcctgcctct aaacagcgtg gaacaatttc acccaagaga
     2761 ccttctgtga ccagtcatgc tgtcaccctc tttagacagt gctccaccat ggcagtgcca
     2821 gacataaaaa ggtcggtctg ggccggagaa accatcacct acagcggatc tcagactgag
     2881 caccatcgcc atggcctgcc ccagtctcgc ttgctgcctg cttggcttac tggctctgac
     2941 ctcggcctgc tacatccaga actgccccct gggcggcaag agggctgtgc tggacctgga
     3001 tatgcgcaag gttagtctcc ccgaccctgt cccttccctt cccgttctgg cgatgctaag
     3061 gaccagagaa gctctcccac ctacagagag cattcccgca cacttgccag ccctaccaag
     3121 gcctcgcgtg ggaacccagg gctttgggaa gtgttaggct ccctcttgac gccgtgaagg
     3181 taacgacaat gccggagcac ccactgcccc tcgctctgcc acagtccgga ttcggattgt
     3241 gcacggcgcc cacccgcatc cttccccaca gtgtctcccc tgcggcccgg gcggcaaagg
     3301 acgctgcttc ggaccaagca tctgctgcgc ggacgagctg ggctgcttcg tgggcaccgc
     3361 cgaggcgctg cgctgccagg aggagaacta cctgccttcg ccctgccagt ctggccagaa
     3421 gccctgcggg agcggaggcc gctgcgccgc cacaggcatc tgctgcagcc cgggtgagca
     3481 ggagggggcc cagcaggtga cccggcaagg agccgtcggg tttgcagctc agaacactga
     3541 cccatttctc ttgcagatgg ctgccgcaca gaccccgcct gcgaccctga gtctgccttc
     3601 tcggagttgc ggcccgggat ccaccggccg gtcgccacca tggtgagcaa gggcgaggag
     3661 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     3721 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     3781 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     3841 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     3901 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     3961 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     4021 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     4081 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     4141 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     4201 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     4261 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     4321 gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcaa gctgagccca
     4381 ctttctggga atacctttag cgcgcttcct tcgttcccca tggccactgc cagaaaaaaa
     4441 aaaaaaaaag aaaagaaaag aaaagaaaag aaaaataaag tagatttcct cttcaaactt
     4501 gactggtgtc taattgtcgg aaacgggagg gaggaaaggc accgggaacg ccgtgctctt
     4561 ggcatcttgt agcaaaaaca gtccagcagg ctgctcgggg accgaggggt gctctatggg
     4621 ctgccatctc ccattagccc catttccatt tcaactgggc tgggcctgat agacttaaga
     4681 gaagaatttg gaagtccaaa cagcaacggt ggacttctgt gggaaagggc aacgcctaac
     4741 aggggactga ggagcctaac aaggcaggct aggaggcagg ggaactagtt cagtcacttc
     4801 catatctgag agggggcttt agctgtgacc tcagaggttg agggggcaat cctggcctag
     4861 aaggcagggt cttggccagg ggagggtagt tacagaacca taaggtttcc acagagctga
     4921 attctcacgc aggccccaag gaaaccacag ctggaaagat caagaaccaa ctggcctctc
     4981 ctctgctgtc cccagggtcg ggcctgagct gggggtgggg gtggggggcg gggggggggc
     5041 tgcccgcctc tgcaggacag cgtgctctgg ggagacatgg gatctaagag ccagtgggca
     5101 caagtgacca caaagcacgg agcccacaga caagcagaaa tccacaatct atactgctcc
     5161 cttactgctg gattaaaata ccgcccttgc cacgaagcgc gcctcacttg gctgccagtg
     5221 ttctcagcag cagtaatggg cagaagtgag ggcatgcccc gcctccacaa gggcacgtga
     5281 ttcgctgtca gaacaaagtg tgagggcttg ggtggagctc agagactgga agtcttgccc
     5341 cacacggtta aagtcctggg ctccatccca tcatagcaaa taagttaaac atacaaacaa
     5401 gcactttctg tggttctccc actctgaaaa tgaacacaga tgccgccccg cccgccccca
     5461 taggtgggat tcacagacag ggtagaggcc aaaaagagac ttcaagattg atttgggttt
     5521 ttctttttct tttttctttt tggtgataat ggggttcttc attttctggg gcttttaaag
     5581 ttagttgttt ttcttttttt aattttatga catttattct actaaatttc catttctgtg
     5641 tacgtgtctc tgtgtgagag tccgtgtaca gtggaccgtg ggcactcttc ccacctcctc
     5701 aacacattta cccctctctg cttgcctggg tttaaccagg aggtacagga gccaggcaga
     5761 ctccaatgcc agcctcctcg tgcagtgccc gcggggacca gaagagggca tcagatcccc
     5821 tgacactgga gttgtgaggt ggttgtgagt catcctaagt gggtgagggg aacagaacgg
     5881 gggggggggg ttctctgcaa gatcagcgtg atcgcctacc tcctgagccg tctccccagc
     5941 ctgagaattg tctaaagtac ttttcactat tgcttcgctg tgtgtttggg ggttgtacgc
     6001 gtgccacagc atctgtgtgg gggtcagagc actcctggca tttgctcaca caagcccctt
     6061 tgcttctaga ggctgcagag aggcggcaag caactactgt gatgtctcgg agtgcccttc
     6121 aatggctctg tagctgagga agggactccc tgaaagggaa gaagaatagc atgtccagta
     6181 ggcacaagta ggctgggcac atgcaactcc tttccttgag aaataggagc ctgattcttc
     6241 cagaaacagg tgttggaaag ggacgtttgc aagtctccac ctgggcagag gagcagagtg
     6301 aggctctcac ctccattctg aaactgtcca tacaccagcc ctagggattg aaccaagtgt
     6361 ggccacacaa cctgccacac ctacctcctc tctgtccctt tttggcctcg cccttccttg
     6421 gccctgccct attggaggtc agaccctgag agggccatgt aaggaggagg ctgggatagg
     6481 agtctgcctg gtccctgtac cccctggtta aacccaggca agcagagagg gggaaaggtg
     6541 ctgaggaggt gggaagagaa ctggagaccg ggtagaagac caagactctg tccagaactg
     6601 accatcctga gccacctagt caggtcactg agactaatct ctttcttccc tgtagttgcc
     6661 cctgacctta gagtctgcag agcatcccaa gaagttatag catagtgtgg ccttaggaga
     6721 ccttgctgca gtcttaggag aagaggctgc ccgggactga tggggcggag agagaaagtg
     6781 ggaagttaac aggagggtga agaacaggga gcccagaaga tggaaggatg gaaaacctga
     6841 gagagcagcc ctctctggcc aaggcaccaa ggggcctgga ggaaggacag gttgggggct
     6901 gaagtgctcg ctgacttggt cgttaaggtc atttaagagg actgggattc agttcatagt
     6961 gcccacatgg taggatgtaa ctacctgtaa ctccaggtcc agggagtctg atgccctctc
     7021 ctgacctctg cagcaccagg cacacacatg atgcccagat atacatacat atataatgaa
     7081 aaagcatcca tatacataaa taaatgaata aatctaaaat tttagaaaaa aaagagaagc
     7141 taatgagaag gggagggaag gagtgccaaa gagcgtggtc tgggcgctac tgtcagcatg
     7201 tgtgcgcgct gctgctgggc tcatgcatct ctgagtcccg agctatggtt gcggtcaaga
     7261 gtcatagaaa cgcccctagc agttgttacc ctttcctcct cttccccttt cccctcctct
     7321 tccatctcca tggtgctggg gaccaaattc aggccaggga tgccttctgc tcctgagaca
     7381 caactccagc ctatcttgtc tatttccgtg cggggagtaa gggcgtggca gtttgaacac
     7441 gtgcctgtaa ggggcttgta gctaggctct cactggggtt tgcctctcag tggttggtgt
     7501 ggagcacctg gagagaatgc aggtggcgtt gtctccttta ggtttggggt aacatctctg
     7561 taccttctcc catccaagca cagtcttggg tgagtggaag tccttcaagc tgtttttgcc
     7621 ttaggtgaaa tcagggctga gatagccacc cctggcttgt cccccccttt ctttgttact
     7681 cttgccgatc tgtctctccc catatccagc tcagttcctt ttgtccacca atggcctgcc
     7741 tctctccttt tattccggtc tcagccaaag aatcagtaag tcaccctgac ccttcagggc
     7801 cgctctgggt cccagacctt tcccctcttc tgagcttggg gttgcctttg tcctgagtat
     7861 taggtgggta tccaggagca cccgatcgct tcctttattc tataagactt acaggttctg
     7921 ccccttctgg tggggaaggg ggcgtggact atgcacgact tcgggtgtgg gggttagcca
     7981 gggcagagta gaaagttccc actccaacgc ctctccctcc cctcttctcc ccaccccaga
     8041 aatagagaca gacaccaggg tgcagttttt aaaagtttat tttccatgct gtaggggcga
     8101 ggcgtggggg cgatggctca gtagacctcg agggggggcc cggtacccaa ttcgccctat
     8161 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     8221 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     8281 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     8341 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     8401 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     8461 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     8521 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     8581 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     8641 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     8701 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     8761 aacgcgaatt ttaacaaaat attaacgctt acaatttag
//
