LOCUS       sequence-131156-        9773 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses the extracellular region of the Neogenin 1, isoform h 
            protein, C-terminally fused to alkaline phosphatase + 6X histidine 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9773)
  AUTHORS   Visser JJ, Cheng Y, Perry SC, Chastain AB, Parsa B, Masri SS, Ray 
            TA, Kay JN, Wojtowicz WM
  TITLE     An extracellular biochemical screen reveals that FLRTs and Unc5s 
            mediate neuronal subtype recognition in the retina.
  JOURNAL   Elife. 2015 Dec 3;4. pii: e08149. doi: 10.7554/eLife.08149.
  PUBMED    26633812
REFERENCE   2  (bases 1 to 9773)
  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: "Elife."; 
            date: "2015-12-3"; volume: "4. pii: e08149. doi"; pages: " 
            10.7554/eLife.08149"
COMMENT     Sequence Label: Neo1.h-AP-His
FEATURES             Location/Qualifiers
     source          1..9773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        25..408
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        409..612
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     563..583
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     609..633
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          748..1573
                     /label=CMV intron A
                     /note="human cytomegalovirus intron A (Chapman et al., 
                     1991)"
     regulatory      1604..1613
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1608..1617
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6353..6370
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    6396..6517
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6433..6452)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6487..6506
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6555..6572)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6726..7314)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6806..6825)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7485..8276,8277..8345))
                     /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     8108..8127
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8346..8450)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8518..8536
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8574..8596)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8598..8618)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8603..8933
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8784..8919
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8846..8865
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     9070..9089
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(9156..9611)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9293..9314)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9505..9524
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9737..9759
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     9751..9768
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     9752..9768
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gtcgaggtct cgacggtatc gattgacatt gattattgac tagctagtta ttaatagtaa
       61 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      121 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      181 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      241 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt
      301 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac
      361 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      421 tggcagtaca ccaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      481 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      541 cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      601 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
      661 gacctccata gaagacaccg ggaccgatcc agcctccgcg gccgggaacg gtgcattgga
      721 acgcggattc cccgtgccaa gagtgacgta agtaccgcct atagactcta taggcacacc
      781 cctttggctc ttatgcatgc tatactgttt ttggcttggg gcctatacac ccccgcttcc
      841 atatgctata ggtgatggta tagcttagcc tataggtgtg ggttattgac cattattgac
      901 cactccccta ttggtgacga tactttccat tactaatcca taacatggct ctttgccaca
      961 actatctcta ttggctatat gccaatactc tgtccttcag agactgacac ggactctgta
     1021 tttttacagg atggggtccc atttattatt tacaaattca catatacaac aacgccgtcc
     1081 cccgtgcccg cagtttttat taaacatagc gtgggatctc cacgcgaatc tcgggtacgt
     1141 gttccggaca tgggctcttc tccggtagcg gcggagcttc cacatccgag ccctggtccc
     1201 atgcctccag cggctcatgg tcgctcggca gctccttgct cctaacagtg gaggccagac
     1261 ttaggcacag cacaatgccc accaccacca gtgtgccgca caaggccgtg gcggtagggt
     1321 atgtgtctga aaatgagctc ggagattggg ctcgcaccgc tgacgcagat ggaagactta
     1381 aggcagcggc agaagaagat gcaggcagct gagttgttgt attctgataa gagtcagagg
     1441 taactcccgt tgcggtgctg ttaacggtgg agggcagtgt agtctgagca gtactcgttg
     1501 ctgccgcgcg cgccaccaga cataatagct gacagactaa cagactgttc ctttccatgg
     1561 gtcttttctg cagaagcttg aattctcgag gatccgcggc cgcgccacca tggcggcgga
     1621 gcgcgaagcc gggcgactcc tctgcacctc ctcctcccgg cgctgctgtc cgccaccgcc
     1681 gctgctgctg ttgctgccgc tgctgctgct gctcggacgc ccggcgtccg gcgccgcggc
     1741 cacgaagagc ggctccccgc cgcagtccgc aggagccagt gttcgaacat tcactccgtt
     1801 ttattttctg gtggagccag tagacaccct ctcagttaga ggctcttctg ttatattaaa
     1861 ttgctcggca tattctgagc cctctccaaa cattgaatgg aagaaagatg ggactttttt
     1921 aaacttagaa tcagatgatc gacgccagct actcccagat ggatctttat tcatcagcaa
     1981 cgtggtgcat tccaaacaca ataagcctga cgaaggtttc tatcagtgtg tagccactgt
     2041 ggataatctt ggaaccattg tcagcagaac agccaagctc acagtagcag gtcttccaag
     2101 atttaccagc caaccagaac cttcttcagt ctatgttgga aacagtgcaa ttctgaattg
     2161 tgaagttaat gcagatttgg tcccatttgt taggtgggaa cagaatcgac agccccttct
     2221 tctagatgac aggattgtca aacttccaag tggaacactg gttatcagca atgctactga
     2281 aggagatggg ggactctacc gctgcattgt tgaaagtggt gggccaccaa agtttagtga
     2341 cgaagctgaa ttgaaagttc ttcaagatcc tgaggaaatt gtagacttgg tatttctgat
     2401 gcgaccatct tctatgatga aagtcactgg tcagagtgca gtgttgccat gtgttgtctc
     2461 agggcttcct gctccagttg ttagatggat gaaaaacgaa gaagtgcttg acacagaaag
     2521 ctctggcagg ttggtcttgc tagcaggagg ttgcttggag atcagtgatg tcactgagga
     2581 tgatgctggg acttattttt gcatagctga taatggaaat aagacagttg aagctcaggc
     2641 ggagcttact gtgcaagtgc cacctggatt cctgaaacaa cctgctaaca tatatgctca
     2701 cgaatccatg gacattgtat ttgaatgtga agtcactggg aagccaactc caactgtgaa
     2761 gtgggtcaag aatggggatg tggttatccc cagtgattac tttaaaattg taaaggaaca
     2821 taatcttcaa gttttgggtc tggtgaaatc agatgaaggg ttctatcaat gcattgctga
     2881 gaatgatgtt ggaaatgcac aagctggagc ccagctgata atccttgagc atgcaccagc
     2941 cacaacggga ccattacctt cagctcctcg agacgtcgtg gcctccctgg tctctactcg
     3001 cttcattaaa ttgacatggc gtacacctgc atcagaccct catggagaca atctcaccta
     3061 ctctgtgttc tacaccaagg aaggggttgc tagggagcgt gttgagaata ccagccagcc
     3121 aggagagatg caggtgacta ttcaaaactt gatgccagca actgtgtaca tcttcaaagt
     3181 tatggctcaa aataagcatg gctctggaga aagttcagct cctcttcgag tagagacaca
     3241 gcctgaggtt cagctccctg gcccagcacc taatatccgt gcttatgcaa cgtcacctac
     3301 ttctatcact gtcacctggg aaacaccgtt atctggcaat ggggaaattc aaaattacaa
     3361 attgtactac atggaaaaag gaactgataa agaacaggat attgatgttt caagtcactc
     3421 ctacaccatt aatggactga agaaatacac agaatacagt ttccgagtgg tggcctacaa
     3481 taaacatggt cctggagttt ctacacaaga tgttgctgtt cgaacattat cagatgttcc
     3541 cagtgctgct cctcagaatc tgtccttaga agtgagaaat tcaaagagta tagtgatcca
     3601 ctggcagccc ccttcctcaa ccacacaaaa tgggcagata actggctaca agattcgata
     3661 tcgaaaggcc tcccgaaaaa gtgatgtcac tgagaccttg gtaactggga cacagctgtc
     3721 tcagctgatt gaaggtcttg atcgggggac agaatataac ttccgagtcg ctgctctcac
     3781 agtcaatggt acaggtccag caactgattg gctgtctgct gaaacttttg aaagcgacct
     3841 agatgaaact cgtgttcctg aagtgcccag ctctcttcat gtccgtccgc tcgtcactag
     3901 cattgtagtg agctggactc ctccagagaa ccagaacatt gtggtccgag gttatgccat
     3961 cggttacggc attggcagcc ctcatgccca gaccatcaaa gtggactata aacaacgtta
     4021 ttacaccatc gaaaacttgg atccaagctc tcattacgtg attaccttga aagcatttaa
     4081 caatgttggc gaaggcatcc ccctttatga gagtgctgtg accagacctc acacagtgcc
     4141 agatcccact cccatgatgc caccagtggg agttcaggct tccattctga gtcacgacac
     4201 cataaggatt acctgggcag acaactccct gcccaaacac cagaagatta cagactcccg
     4261 ctactacaca gtccggtgga agaccaacat cccagcaaac acgaagtaca agaatgcaaa
     4321 tgcaacgacg ttaagctatt tggttactgg tttaaagcca aatacgctct atgagttctc
     4381 tgtgatggtg accaaaggca gaaggtcaag cacgtggagt atgacagctc atggcgctac
     4441 ctttgaatta gttcctactt ctccacctaa ggatgtgaca gttgtgagta aggaaggaaa
     4501 acctagaacc atcatagtga actggcagcc tccctctgaa gctaacggca agattacagg
     4561 ttacatcatc tattacagca cggatgtgaa tgcagagata catgactggg ttattgaacc
     4621 agttgtggga aacagactga ctcaccagat tcaagagtta acacttgata cgccatacta
     4681 cttcaaaatc caggcccgga actcaaaggg catggggccc atgtctgaag ctgtacagtt
     4741 cagaacacct aaagccttag ggtcagcagg aaaaggaagc cgactaccag acctgggatc
     4801 tgactacaaa cctccaatga gtggcagcaa cagccctcac gggagcccca cctcccctct
     4861 ggacagcact agtggatctt ccggaatcat cccagttgag gaggagaacc cggacttctg
     4921 gaaccgcgag gcagccgagg ccctgggtgc cgccaagaag ctgcagcctg cacagacagc
     4981 cgccaagaac ctcatcatct tcctgggcga tgggatgggg gtgtctacgg tgacagctgc
     5041 caggatccta aaagggcaga agaaggacaa actggggcct gagatacccc tggccatgga
     5101 ccgcttccca tatgtggctc tgtccaagac atacaatgta gacaaacatg tgccagacag
     5161 tggagccaca gccacggcct acctgtgcgg ggtcaagggc aacttccaga ccattggctt
     5221 gagtgcagcc gcccgcttta accagtgcaa cacgacacgc ggcaacgagg tcatctccgt
     5281 gatgaatcgg gccaagaaag cagggaagtc agtgggagtg gtaaccacca cacgagtgca
     5341 gcacgcctcg ccagccggca cctacgccca cacggtgaac cgcaactggt actcggacgc
     5401 cgacgtgcct gcctcggccc gccaggaggg gtgccaggac atcgctacgc agctcatctc
     5461 caacatggac attgacgtga tcctaggtgg aggccgaaag tacatgtttc gcatgggaac
     5521 cccagaccct gagtacccag atgactacag ccaaggtggg accaggctgg acgggaagaa
     5581 tctggtgcag gaatggctgg cgaagcgcca gggtgcccgg tatgtgtgga accgcactga
     5641 gctcatgcag gcttccctgg acccgtctgt gacccatctc atgggtctct ttgagcctgg
     5701 agacatgaaa tacgagatcc accgagactc cacactggac ccctccctga tggagatgac
     5761 agaggctgcc ctgcgcctgc tgagcaggaa cccccgcggc ttcttcctct tcgtggaggg
     5821 tggtcgcatc gaccatggtc atcatgaaag cagggcttac cgggcactga ctgagacgat
     5881 catgttcgac gacgccattg agagggcggg ccagctcacc agcgaggagg acacgctgag
     5941 cctcgtcact gccgaccact cccacgtctt ctccttcgga ggctaccccc tgcgagggag
     6001 ctccatcttc gggctggccc ctggcaaggc ccgggacagg aaggcctaca cggtcctcct
     6061 atacggaaac ggtccaggct atgtgctcaa ggacggcgcc cggccggatg ttaccgagag
     6121 cgagagcggg agccccgagt atcggcagca gtcagcagtg cccctggacg aagagaccca
     6181 cgcaggcgag gacgtggcgg tgttcgcgcg cggcccgcag gcgcacctgg ttcacggcgt
     6241 gcaggagcag accttcatag cgcacgtcat ggccttcgcc gcctgcctgg agccctacac
     6301 cgcctgcgac ctggcgcccc ccgccggcac caccgacgcc gcgcacccgg gtcatcatca
     6361 ccatcaccat taatctagag atcttattaa agcagaactt gtttattgca gcttataatg
     6421 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6481 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggtcg actctagcta
     6541 gactcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     6601 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     6661 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6721 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6781 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6841 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6901 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6961 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     7021 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     7081 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     7141 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     7201 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     7261 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     7321 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     7381 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     7441 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     7501 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     7561 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     7621 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     7681 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     7741 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     7801 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     7861 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     7921 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     7981 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     8041 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     8101 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     8161 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     8221 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     8281 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     8341 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     8401 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     8461 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     8521 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     8581 cacatgcagc tcccggagga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     8641 aggcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     8701 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     8761 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     8821 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     8881 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     8941 cgggagcttg tatatccatt ttcgtcccgg agacggtcac agcttgtctg taagcggatg
     9001 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     9061 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     9121 cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt
     9181 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     9241 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     9301 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     9361 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     9421 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     9481 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     9541 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     9601 acagggcgcg tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     9661 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     9721 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga att
//
