LOCUS       PAL-AA245-428-GF        5279 bp DNA     circular SYN 04-SEP-2025
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5279)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 4, 2025 from SnapGene 7.1.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5279
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    591..646
                     /label=MCS
                     /note="multiple cloning site of fluorescent protein
                     plasmids"
                     /note="color: #99ccff"
     primer_bind     646..673
                     /label=Primer 1
                     /note="color: black; sequence: 
                     GGACTCGAATTCGGTCTGGCTGAGTTGGACCCAG; added: 2025-09-04"
     misc_feature    join(652..1206,1207..1217)
                     /label=MCS
                     /note="multiple cloning site of fluorescent protein
                     plasmids"
                     /note="This feature has 2 segments:
                        1:  652 .. 1206 / #99ccff
                        2: 1207 .. 1217 / #99ccff"
     CDS             652..1203
                     /label=Arl13B CDS
                     /label=Arl13B\CDS
                     /note="/vntifkey=21"
                     /note="color: #ff6600; direction: RIGHT"
     primer_bind     complement(1181..1211)
                     /label=Primer 2
                     /note="color: black; sequence: 
                     CTACGGATCCAATGAGATCACATCATGAGCATCAC; added: 2025-09-04"
     CDS             join(1225..1227,1228..1230,1231..1944)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1: 1225 .. 1227 / #05fd14
                        2: 1228 .. 1230 / #05fd14 / 1a
                        3: 1231 .. 1944 / #05fd14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    2067..2188
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      complement(2195..2650)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        2677..2781
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     promoter        2783..3140
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      2991..3126
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             3175..3969
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin)"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    4201..4248
                     /label=HSV TK poly(A) signal
                     /note="herpesvirus thymidine kinase polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      4577..5165
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagat gttcagtctc atggcaagct gttgtgaatt cggtctggct
      661 gagttggacc cagaaccaac gaatcctttc cagccaatag catctgtaat cattgagaat
      721 gaaggaaaac ttgaaagaga gaaaaaaaac caaaaaatgg agaaagacag tgatggctgc
      781 cacctgaaac ataaaatgga gcatgagcaa atagagacac aaggccaggt taatcacaat
      841 ggccaaaaaa ataatgaatt tggactagta gaaaattata aggaggcatt aacacagcag
      901 ttaaagaatg aagatgagac agaccggcca tcattggaat cagctaatgg taaaaagaaa
      961 actaagaaac taagaatgaa aaggaaccac cgggtagaac cacttaatat agatgactgt
     1021 gctcctgaga gtccaacgcc acccccaccc cctcctcctg ttggctgggg aacccctaaa
     1081 gtcactagac ttccaaaact tgagcctctt ggtgaaacac atcataatga tttctatagg
     1141 aagccactgc ctcccctggc tgtgccacag cgacctaaca gtgatgctca tgatgtgatc
     1201 tcattggatc caccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
     1261 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     1321 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     1381 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     1441 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     1501 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     1561 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1621 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     1681 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     1741 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1801 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     1861 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     1921 ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca
     1981 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     2041 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     2101 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     2161 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     2221 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     2281 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     2341 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     2401 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     2461 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     2521 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     2581 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     2641 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2701 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2761 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     2821 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2881 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2941 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3001 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3061 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3121 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3181 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3241 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3301 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     3361 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3421 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3481 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3541 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3601 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3661 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     3721 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3781 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     3841 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     3901 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     3961 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4021 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4081 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4141 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     4201 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     4261 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     4321 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     4381 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     4441 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4501 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4561 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4621 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4681 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     4741 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4801 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4861 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4921 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4981 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5041 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5101 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5161 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5221 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
