LOCUS       sequence-229563-        5299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For analysis of AIB cell differentiation by tracking Venus 
            fluorescence under the expression of Pinx-1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5299)
  AUTHORS   Hori S, Oda S, Suehiro Y, Iino Y, Mitani S
  TITLE     OFF-responses of interneurons optimize avoidance behaviors depending
            on stimulus strength via electrical synapses.
  JOURNAL   PLoS Genet. 2018 Jun 25;14(6):e1007477. doi: 
            10.1371/journal.pgen.1007477. eCollection 2018 Jun.
  PUBMED    29939997
REFERENCE   2  (bases 1 to 5299)
  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 
            Genet."; date: "2018-06-25"; volume: "14(6):e1007477. doi"; pages: "
            10.1371/journal.pgen.1007477. eCollection 2018 Jun"
COMMENT     Sequence Label: pPD_Pinx-1(1k + 50 bp)::Venus
FEATURES             Location/Qualifiers
     source          1..5299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    105..126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(141..146,147..164,165..171)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    179..195
                     /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     184..206
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     203..219
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     203..219
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(1452..1454,1455..1457,1458..2171)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1497..1518)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1758..1777)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2105..2126
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3087..3109)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3209..3228
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3306..3324)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3392..3496
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3497..3565,3566..4357)
                     /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(3715..3734)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4528..5116
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5017..5036
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5270..5287
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
       61 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
      121 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      181 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctgtaa
      241 gtttaaacat gatcttacta actaactatt ctcatttaaa ttttcagagc ttaaaaatgg
      301 ctgaaatcac tcacaacgat ggatacgcta acaacttgga aatgaaataa gcttgcatgc
      361 ctgcaggtcg acctaaatag attcaaaaat tattttctgt gcttttcaca aatacacttt
      421 attttaggta atcatatttt aattaaacac gcgggaaatt ttttagggta aactttgaag
      481 ttttttttgt cgcggatatc agattcataa ctgtctatgt accataaatt acgacttttg
      541 aatatttgct gttcaaagtc aatgtataaa ttaaatttaa gtttctttcg tcatacatct
      601 caaaaatagc taaaatgcgt ttaacaaatg ttatacctaa ttttcaatca taacttggag
      661 agtatgaatg aattcaaacg tttccttgag gtgatatcct tttggtactt gtatattatc
      721 gaggcacagt ttgaaaataa attaaatttt aattcatttg atgattttgt tttctcttga
      781 ggcttaaaaa tgataaacgg tacaaaacta caaaaaaact ccataagtct ttattttctt
      841 aatttttgaa attttatttc aaaatgcaca agatccattt atcatattta tagttcttca
      901 tcttcttttt tttaatatcc tctttttgtg ttttattcgg tccatacgtg ctcttcaact
      961 tttttttcat ttttgacttt aaataagaaa agatggcaat aaaaaatttg gccgaagagg
     1021 gcgatggacg gatgaaaatc tactaaaagg attataactc aatttgatat gctcttcggg
     1081 aggtatcctc ctgacgagat ggaaaagaag aagaagaaga agaagaagct tgtcatcgtt
     1141 tcatcggcaa agagacgggt ggacattaga ccacgcccca cagggaaacc tttcgagttt
     1201 catccacctc tgtctgttta acatatttgt ttttttctag ctctattttt cttccgcttc
     1261 tactgcgtac ttttgcataa ttctatttct aagtctgatc attataagta tcatcctgaa
     1321 catcgcacac taaacatcct cggccggcac ggatgaagta gttttcattg cagttcttgt
     1381 ccgccggaat gcttctatat tatctggcgg ccatattcaa gggcttacat ccgcgagtgg
     1441 atccccgggc tatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1501 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1561 atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc
     1621 cctggcccac cctcgtgacc accctgggct acggcctgca gtgcttcgcc cgctaccccg
     1681 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1741 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     1801 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1861 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca
     1921 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg
     1981 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     2041 ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac gagaagcgcg
     2101 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     2161 tgtacaagta agaattccaa ctgagcgccg gtcgctacca ttaccaactt gtctggtgtc
     2221 aaaaataata ggggccgctg tcatcagagt aagtttaaac tgagttctac taactaacga
     2281 gtaatattta aattttcagc atctcgcgcc cgtgcctctg acttctaagt ccaattactc
     2341 ttcaacatcc ctacatgctc tttctccctg tgctcccacc ccctattttt gttattatca
     2401 aaaaacttct cttaatttct ttgtttttta gcttctttta agtcacctct aacaatgaaa
     2461 ttgtgtagat tcaaaaatag aattaattcg taataaaaag tcgaaaaaaa ttgtgctccc
     2521 tccccccatt aataataatt ctatcccaaa atctacacaa tgttctgtgt acacttctta
     2581 tgttttttac ttctgataaa tttttttgaa acatcataga aaaaaccgca cacaaaatac
     2641 cttatcatat gttacgtttc agtttatgac cgcaattttt atttcttcgc acgtctgggc
     2701 ctctcatgac gtcaaatcat gctcatcgtg aaaaagtttt ggagtatttt tggaattttt
     2761 caatcaagtg aaagtttatg aaattaattt tcctgctttt gctttttggg gtttccccta
     2821 ttgtttgtca agatttcgag gacggcgttt ttcttgctaa aatcacaagt attgatgagc
     2881 acgatgcaag aaagatcgga agaaggtttg ggtttgaggc tcagtggaag gtgagtagaa
     2941 gttgataatt tgaaagtgga gtagtgtcta tggggttttt gccttaaatg acagaataca
     3001 ttcccaatat accaaacata actgtttcct actagtcggc cgtacgggcc ctttcgtctc
     3061 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     3121 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     3181 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     3241 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcggcct
     3301 taagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     3361 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3421 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3481 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3541 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3601 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3661 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     3721 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     3781 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     3841 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     3901 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     3961 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4021 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     4081 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4141 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4201 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4261 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4321 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     4381 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4441 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4501 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4561 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4621 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4681 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4741 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4801 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4861 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4921 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4981 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5041 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5101 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5161 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     5221 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     5281 gagcgaggaa gcggaagag
//
