LOCUS       sequence-221637-        5299 bp    DNA     circular SYN 15-APR-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 Apr 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"
     CDS             join(1104..1106,1107..1109,1110..1823)
                     /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(1149..1170)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1410..1429)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1757..1778
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2739..2761)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2861..2880
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2958..2976)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3044..3148
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3149..3217,3218..4009)
                     /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(3367..3386)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4180..4768
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4669..4688
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4922..4939
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5056..5077
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5092..5097,5098..5115,5116..5122)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5130..5146
                     /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     5135..5157
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5154..5170
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5154..5170
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgacctaaat agattcaaaa attattttct gtgcttttca
       61 caaatacact ttattttagg taatcatatt ttaattaaac acgcgggaaa ttttttaggg
      121 taaactttga agtttttttt gtcgcggata tcagattcat aactgtctat gtaccataaa
      181 ttacgacttt tgaatatttg ctgttcaaag tcaatgtata aattaaattt aagtttcttt
      241 cgtcatacat ctcaaaaata gctaaaatgc gtttaacaaa tgttatacct aattttcaat
      301 cataacttgg agagtgtgaa tgaattcaaa cgtttccttg aggtgatatc cttttggtac
      361 ttgtatatta tcgaggcaca gtttgaaaat aaattaaatt ttaattcatt tgatgatttt
      421 gttttctctt gaggcttaaa aatgataaac ggtacaaaac tacaaaaaaa ctccataagt
      481 ctttattttc ttaatttttg aaattttatt tcaaaatgca caagatccat ttatcatatt
      541 tatagttctt catcttcttt tttttaatat cctctttttg tgttttattc ggtccatacg
      601 tgctcttcaa cttttttttc atttttgact ttaaataaga aaagatggca ataaaaaatt
      661 tggccgaaga gggcgatgga cggatgaaaa tctactaaaa ggattataac tcaatttgat
      721 atgctcttcg ggaggtatcc tcctgacgag atggaaaaga agaagaagaa gaagaagaag
      781 cttgtcatcg tttcatcggc aaagagacgg gtggacatta gaccacgccc cacagggaaa
      841 cctttcgagt ttcatccacc tctgtctgtt taacatattt gtttttttct agctctattt
      901 ttcttccgct tctactgcgt acttttgcat aattctattt ctaagtctga tcattataag
      961 tatcatcctg aacatcgcac actaaacatc ctcggccggc acggatgaag tagttttcat
     1021 tgcagttctt gtccgccgga atgcttctat attatctggc ggccatattc aagggcttac
     1081 atccgcgagt ggatccccgg gctatggtga gcaagggcga ggagctgttc accggggtgg
     1141 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1201 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc accaccggca
     1261 agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcctg cagtgcttcg
     1321 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1381 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1441 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1501 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     1561 tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg
     1621 aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1681 ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca
     1741 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1801 gcatggacga gctgtacaag taagaattcc aactgagcgc cggtcgctac cattaccaac
     1861 ttgtctggtg tcaaaaataa taggggccgc tgtcatcaga gtaagtttaa actgagttct
     1921 actaactaac gagtaatatt taaattttca gcatctcgcg cccgtgcctc tgacttctaa
     1981 gtccaattac tcttcaacat ccctacatgc tctttctccc tgtgctccca ccccctattt
     2041 ttgttattat caaaaaactt ctcttaattt ctttgttttt tagcttcttt taagtcacct
     2101 ctaacaatga aattgtgtag attcaaaaat agaattaatt cgtaataaaa agtcgaaaaa
     2161 aattgtgctc cctcccccca ttaataataa ttctatccca aaatctacac aatgttctgt
     2221 gtacacttct tatgtttttt acttctgata aatttttttg aaacatcata gaaaaaaccg
     2281 cacacaaaat accttatcat atgttacgtt tcagtttatg accgcaattt ttatttcttc
     2341 gcacgtctgg gcctctcatg acgtcaaatc atgctcatcg tgaaaaagtt ttggagtatt
     2401 tttggaattt ttcaatcaag tgaaagttta tgaaattaat tttcctgctt ttgctttttg
     2461 gggtttcccc tattgtttgt caagatttcg aggacggcgt ttttcttgct aaaatcacaa
     2521 gtattgatga gcacgatgca agaaagatcg gaagaaggtt tgggtttgag gctcagtgga
     2581 aggtgagtag aagttgataa tttgaaagtg gagtagtgtc tatggggttt ttgccttaaa
     2641 tgacagaata cattcccaat ataccaaaca taactgtttc ctactagtcg gccgtacggg
     2701 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     2761 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     2821 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     2881 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     2941 atcaggcggc cttaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     3001 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3061 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3121 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     3181 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3241 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3301 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3361 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     3421 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3481 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3541 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3601 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     3661 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     3721 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     3781 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     3841 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     3901 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     3961 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4021 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4081 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4141 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4201 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4261 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4321 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4381 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4441 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4501 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4561 acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4621 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4681 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4741 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4801 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     4861 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     4921 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     4981 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     5041 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     5101 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     5161 cagctatgac catgattacg ccaagctgta agtttaaaca tgatcttact aactaactat
     5221 tctcatttaa attttcagag cttaaaaatg gctgaaatca ctcacaacga tggatacgct
     5281 aacaacttgg aaatgaaat
//
