LOCUS       sequence-135599-        8630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the entire Contactin 2, isoform 2 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 8630)
  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 8630)
  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: "Elife."; 
            date: "2015-12-3"; volume: "4. pii: e08149. doi"; pages: " 
            10.7554/eLife.08149"
COMMENT     Sequence Label: Cntn4.2-AP-His
FEATURES             Location/Qualifiers
     source          1..8630
                     /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)"
     CDS             5210..5227
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5253..5374
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5290..5309)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5344..5363
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5412..5429)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5583..6171)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5663..5682)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6342..7133,7134..7202))
                     /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     6965..6984
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7203..7307)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7375..7393
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7431..7453)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7455..7475)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7460..7790
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7641..7776
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7703..7722
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7927..7946
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(8013..8468)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8150..8171)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8362..8381
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8594..8616
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8608..8625
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8609..8625
                     /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 tgaggttgcc
     1621 atgggaactg ctagtgctgc aatcattcat gttgtgcctt gcagatgact acacactgca
     1681 tggcccagtt tttgttcaag aaccaagtca tgtcatgttt cctttggatt ctgaggagaa
     1741 aaaagtgaaa ctcagttgtg aagttaaagg gaatccaaaa cctcacatca ggtggaagtt
     1801 aaatggaaca gatgttgaca ttggtatgga tttccgctac agtgttgttg acggcagctt
     1861 gttgatcaat aaccccaata aaacccaaga tgctggaacg taccagtgca tagcaacaaa
     1921 ctcatttgga acaattgtaa gcagagaagc aaagcttcag tttgcttatc ttgaaaactt
     1981 taagacaaga acaagaagca ctgtgtcagt ccgtcgaggt cagggaatgg tccttctgtg
     2041 cggcccgcca ccccattctg gagagctgag ctatgcctgg atcttcaatg aatacccttc
     2101 ctatcaggat aaccgccgct tcgtttctca agagactggg aatctgtata ttgccaaagt
     2161 ggaaaaatct gatgttggta attacacctg tgtagttacc aacactgtga ccaaccacaa
     2221 ggtcctggga ccgcccacac cactaatcct gagaaatgat ggggtgatgg gtgaatatga
     2281 gcccaaaata gaagtacagt ttccagaaac cgtcccggct gagaaaggaa caacagtcaa
     2341 actggaatgc tttgccttag gaaatccagt cccaactatt ctatggcgaa gagctgatgg
     2401 caagccaata gcaaggaaag ccagaagaca taagtcgaat ggaattctgg aaattcccaa
     2461 ttttcagcag gaagatgccg gttcatatga atgtgtggct gaaaactcca gaggcaaaaa
     2521 cgtagccaag ggacagttga ctttttatgc tcaacctaac tgggttcaaa taataaatga
     2581 tatccacgtg gccatggaag agagtgtctt ctgggaatgt aaagcaaatg ggaggcctaa
     2641 acctacctac agatggctaa aaaatggtga cccactgtta acacgggata gaatacaaat
     2701 cgagcaagga acactcaaca taacaatagt gaatctctca gatgctggca tgtaccagtg
     2761 tgtggcagaa aataaacatg gagttatctt ttccagcgct gagcttagtg taatagctga
     2821 aagtccagat ttttcccgaa cactcttaaa aagagtaaca cttgtcaaag tgggaggtga
     2881 agttgtcatt gagtgcaagc caaaagcatc cccaagacct gtctacacct ggagaaaggg
     2941 aagagaaata ttaagagaaa atgaaagaat taccatttct gaagatggga acctcagaat
     3001 catcaatgtg actaaatctg atgctggaag ttacacctgc atagccacga accattttgg
     3061 aactgccagc agtactggga atgtggttgt gaaagaccca acaaaggtga tggttcctcc
     3121 ttccagcatg gatgtcactg taggagaaag cattgttctc ccatgccagg taacacatga
     3181 tcactccctg gacattgtgt ttacgtggac atttaatgga cacttgatag actttgacaa
     3241 ggatggagac cactttgaaa gagttggagg gcaggattca gctggtgatt tgatgatccg
     3301 aaacatccaa ctgaagcatg ctgggaagta tgtctgcatg gtccaaacaa gtgtagacaa
     3361 actctctgtt gctgcggacc tgattgtaag aggtcctcca ggtcctcctg aggctgtgac
     3421 catagatgaa atcacagaca ccactgctca gctttcttgg agaccaggac ctgacaacca
     3481 cagccccatc accatgtatg ttatccaagc taggactcca ttctccgtgg gctggcaagc
     3541 agtcaataca gttccagatc tcgtcgacgg gaagacattc acagcaactg tggtgggttt
     3601 gaatccttgg gttgaatatg aattccgcac tgttgcagct aatgtgattg gcattgggga
     3661 acccagccgg ccctcagaga aacggaggac agaagaggct cagaacactg gggatcagag
     3721 aggtactagt ggatcttccg gaatcatccc agttgaggag gagaacccgg acttctggaa
     3781 ccgcgaggca gccgaggccc tgggtgccgc caagaagctg cagcctgcac agacagccgc
     3841 caagaacctc atcatcttcc tgggcgatgg gatgggggtg tctacggtga cagctgccag
     3901 gatcctaaaa gggcagaaga aggacaaact ggggcctgag atacccctgg ccatggaccg
     3961 cttcccatat gtggctctgt ccaagacata caatgtagac aaacatgtgc cagacagtgg
     4021 agccacagcc acggcctacc tgtgcggggt caagggcaac ttccagacca ttggcttgag
     4081 tgcagccgcc cgctttaacc agtgcaacac gacacgcggc aacgaggtca tctccgtgat
     4141 gaatcgggcc aagaaagcag ggaagtcagt gggagtggta accaccacac gagtgcagca
     4201 cgcctcgcca gccggcacct acgcccacac ggtgaaccgc aactggtact cggacgccga
     4261 cgtgcctgcc tcggcccgcc aggaggggtg ccaggacatc gctacgcagc tcatctccaa
     4321 catggacatt gacgtgatcc taggtggggg ccgaaagtac atgtttcgca tgggaacccc
     4381 agaccctgag tacccagatg actacagcca aggtgggacc aggctggacg ggaagaatct
     4441 ggtgcaggaa tggctggcga agcgccaggg tgcccggtat gtgtggaacc gcactgagct
     4501 catgcaggct tccctggacc cgtctgtgac ccatctcatg ggtctctttg agcctggaga
     4561 catgaaatac gagatccacc gagactccac actggacccc tccctgatgg agatgacaga
     4621 ggctgccctg cgcctgctga gcaggaaccc ccgcggcttc ttcctcttcg tggagggtgg
     4681 tcgcatcgac catggtcatc atgaaagcag ggcttaccgg gcactgactg agacgatcat
     4741 gttcgacgac gccattgaga gggcgggcca gctcaccagc gaggaggaca cgctgagcct
     4801 cgtcactgcc gaccactccc acgtcttctc cttcggaggc taccccctgc gagggagctc
     4861 catcttcggg ctggcccctg gcaaggcccg ggacaggaag gcctacacgg tcctcctata
     4921 cggaaacggt ccaggctatg tgctcaagga cggcgcccgg ccggatgtta ccgagagcga
     4981 gagcgggagc cccgagtatc ggcagcagtc agcagtgccc ctggacgaag agacccacgc
     5041 aggcgaggac gtggcggtgt tcgcgcgcgg cccgcaggcg cacctggttc acggcgtgca
     5101 ggagcagacc ttcatagcgc acgtcatggc cttcgccgcc tgcctggagc cctacaccgc
     5161 ctgcgacctg gcgccccccg ccggcaccac cgacgccgcg cacccgggtc atcatcacca
     5221 tcaccattaa tctagagatc ttattaaagc agaacttgtt tattgcagct tataatggtt
     5281 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5341 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggtcgact ctagctagac
     5401 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5461 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5521 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5581 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5641 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5701 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5761 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5821 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5881 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5941 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6001 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     6061 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6121 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6181 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6241 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6301 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6361 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6421 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6481 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6541 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6601 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6661 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6721 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6781 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6841 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6901 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6961 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7021 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7081 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7141 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7201 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7261 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7321 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     7381 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     7441 atgcagctcc cggaggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagg
     7501 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     7561 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     7621 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     7681 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     7741 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     7801 gagcttgtat atccattttc gtcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     7861 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     7921 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     7981 acagatgcgt aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa
     8041 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     8101 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     8161 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     8221 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     8281 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     8341 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     8401 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     8461 gggcgcgtcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     8521 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     8581 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
//
