LOCUS       sequence-151433-        6770 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tracing neuronal projection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6770)
  AUTHORS   Delwig A, Larsen DD, Yasumura D, Yang CF, Shah NM, Copenhagen DR
  TITLE     Retinofugal Projections from Melanopsin-Expressing Retinal Ganglion 
            Cells Revealed by Intraocular Injections of Cre-Dependent Virus.
  JOURNAL   PLoS One. 2016 Feb 19;11(2):e0149501. doi: 
            10.1371/journal.pone.0149501. eCollection 2016.
  PUBMED    26895233
REFERENCE   2  (bases 1 to 6770)
  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 One.";
            date: "2016-02-19"; volume: "11(2):e0149501. doi"; pages: " 
            10.1371/journal.pone.0149501. eCollection 2016"
COMMENT     Sequence Label: AAV-flex-plap
FEATURES             Location/Qualifiers
     source          1..6770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     enhancer        217..520
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        521..724
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     675..695
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     721..745
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          873..1251
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     protein_bind    complement(1358..1391)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(1402..1435)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             complement(1581..3086)
                     /codon_start=1
                     /product="secreted alkaline phosphatase from human 
                     placenta"
                     /label=SEAP
                     /note="The natural enzyme has a C-terminal transmembrane 
                     sequence that is replaced by a GPI anchor."
                     /translation="MLLLLLLLLGLRLQLSLGIIPVEEENPDFWNREAAEALGAAKKLQ
                     PAQTAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKTYNVD
                     KHVPDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGKSVGV
                     VTTTRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVILGGGR
                     KYMFRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLAKRQGARYVWNRTELMQASLDPSVT
                     HLMGLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRNPRGFFLFVEGGRIDHGHHES
                     RAYRALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSIFGLAPGK
                     ARDRKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGEDVAVF
                     ARGPQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTD"
     protein_bind    3111..3144
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3155..3188
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     misc_feature    3198..3786
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3251..3271)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3669..3680)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3797..3814)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3803..4010
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4033..4173
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4033..4162
                     /label=AAV2 ITR
     rep_origin      4248..4703
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4335..4354)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4545..4566
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4720..4739)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4839..4861
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4899..4917)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4985..5089
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5090..5158,5159..5950)
                     /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(5308..5327)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6121..6709
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6610..6629
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtctagt tattaatagt aatcaattac
      181 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      241 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      301 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      361 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      421 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      481 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      541 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      601 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      661 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      721 agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca
      781 tagaagacac cgggaccgat ccagcctccg cggattcgaa tcccggccgg gaacggtgca
      841 ttggaacgcg gattccccgt gccaagagtg acgtaagtac cgcctataga gtctataggc
      901 ccacaaaaaa tgctttcttc ttttaatata cttttttgtt tatcttattt ctaatacttt
      961 ccctaatctc tttctttcag ggcaataatg atacaatgta tcatgcctct ttgcaccatt
     1021 ctaaagaata acagtgataa tttctgggtt aaggcaatag caatatttct gcatataaat
     1081 atttctgcat ataaattgta actgatgtaa gaggtttcat attgctaata gcagctacaa
     1141 tccagctacc attctgcttt tattttatgg ttgggataag gctggattat tctgagtcca
     1201 agctaggccc ttttgctaat catgttcata cctcttatct tcctcccaca gctcctgggc
     1261 aacgtgctgg tctgtgtgct ggcccatcac tttggcaaag aattgggatt cgaacatcga
     1321 tagcgacgcg tggacggtac ccttcaggtc gacgtcgata acttcgtata gcatacatta
     1381 tacgaagtta tccgaatcgc aataacttcg tataggatac tttatacgaa gttatttggc
     1441 gcgccaaatt taaataacgc aacgtacctt cgagggggcc caaggaattc tcagggagca
     1501 gtggccgtct ccagcagcag cagggtcccg gccagcagag gaagcaacgc ggggaccacg
     1561 gaccgccccg ggtgcgcggc gtcggtggtg ccggcggggg gcgccaggtc gcaggcggtg
     1621 tagggctcca ggcaggcggc gaaggccatg acgtgcgcta tgaaggtctg ctcctgcacg
     1681 ccgtgaacca ggtgcgcctg cgggccgcgc gcgaacaccg ccacgtcctc gcctgcgtgg
     1741 gtctcttcgt ccaggggcac tgctgactgc tgccgatact cggggctccc gctctcgctc
     1801 tcggtaacat ccggccgggc gccgtccttg agcacatagc ctggaccgtt tccgtatagg
     1861 aggaccgtgt aggccttcct gtcccgggcc ttgccagggg ccagcccgaa gatggagctc
     1921 cctcgcaggg ggtagcctcc gaaggagaag acgtgggagt ggtcggcagt gacgaggctc
     1981 agcgtgtcct cctcgctggt gagctggccc gccctctcaa tggcgtcgtc gaacatgatc
     2041 gtctcagtca gtgcccggta agccctgctt tcatgatgac catggtcgat gcgaccaccc
     2101 tccacgaaga ggaagaagcc gcgggggttc ctgctcagca ggcgcagggc agcctctgtc
     2161 atctccatca gggaggggtc cagtgtggag tctcggtgga tctcgtattt catgtctcca
     2221 ggctcaaaga gacccatgag atgggtcaca gacgggtcca gggaagcctg catgagctca
     2281 gtgcggttcc acacataccg ggcaccctgg cgcttcgcca gccattcctg caccagattc
     2341 ttcccgtcca gcctggtccc accttggctg tagtcatctg ggtactcagg gtctggggtt
     2401 cccatgcgaa acatgtactt tcggcctcca cctaggatca cgtcaatgtc catgttggag
     2461 atgagctgcg tagcgatgtc ctggcacccc tcctggcggg ccgaggcagg cacgtcggcg
     2521 tccgagtacc agttgcggtt caccgtgtgg gcgtaggtgc cggctggcga ggcgtgctgc
     2581 actcgtgtgg tggttaccac tcccactgac ttccctgctt tcttggcccg attcatcacg
     2641 gagatgacct cgttgccgcg tgtcgtgttg cactggttaa agcgggcggc tgcactcaag
     2701 ccaatggtct ggaagttgcc cttgaccccg cacaggtagg ccgtggctgt ggctccactg
     2761 tctggcacat gtttgtctac attgtatgtc ttggacagag ccacatatgg gaagcggtcc
     2821 atggccaggg gtatctcagg ccccagtttg tccttcttct gcccttttag gatcctggca
     2881 gctgtcaccg tagacacccc catcccatcg cccaggaaga tgatgaggtt cttggcggct
     2941 gtctgtgcag gctgcagctt cttggcggca cccagggcct cggctgcctc gcggttccag
     3001 aagtccgggt tctcctcctc aactgggatg atgcccaggg agagctgtag cctcaggccc
     3061 agcagcagca gcagcagcag cagcatggtg gcaaggccgg cctcgagttg ataacttcgt
     3121 ataatgtatg ctatacgaag ttatccgaat cgcaataact tcgtataaag tatcctatac
     3181 gaagttatcg aaagcttaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     3241 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     3301 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     3361 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     3421 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     3481 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     3541 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     3601 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     3661 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     3721 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     3781 ccccgcctga ccggtgcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     3841 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     3901 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     3961 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga cacgtgcgga
     4021 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     4081 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     4141 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     4201 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     4261 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4321 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4381 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4441 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     4501 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4561 aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc
     4621 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     4681 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     4741 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     4801 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     4861 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     4921 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     4981 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5041 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     5101 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     5161 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     5221 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     5281 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     5341 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     5401 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5461 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     5521 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5581 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     5641 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5701 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5761 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5821 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5881 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5941 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     6001 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6061 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6121 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6181 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6241 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     6301 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6361 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6421 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6481 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6541 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6601 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6661 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6721 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
