LOCUS       pNeonGreen-C1-Ni        5614 bp    DNA     circular SYN 13-OCT-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5614)
  AUTHORS   Claire Weckerly
  TITLE     Direct Submission
  JOURNAL   Exported Oct 13, 2025 from SnapGene 8.2.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5614
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..589
                     /label=CMV
                     /note="CMV"
     misc_feature    1
                     /label=Kanamycin
                     /note="Kanamycin"
     primer_bind     519..539
                     /label=CMV-F
     CDS             613..1320
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     primer_bind     613..630
                     /label=FPi-F
                     /note="Amplify fluorescent protein from eGFP/CFP etc pr 
                     mCherry etc"
     primer_bind     616..633
                     /label=FPi-F2.0
     primer_bind     complement(616..633)
                     /label=FPv-R
                     /note="Amplify vector without eGFP/CFP etc pr mCherry etc. 
                     Use to exchange with FPi primers."
     regulatory      868..877
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1202..1222
                     /label=NeonGFWD1
     primer_bind     complement(1301..1335)
                     /label=NG-NIR1-VR2
     primer_bind     1301..1320
                     /label=FPv-F
                     /note="Amplify vector without eGFP/CFP etc pr mCherry etc. 
                     Use to exchange with FPi primers."
     primer_bind     complement(1301..1320)
                     /label=FPi-R
                     /note="Amplify fluorescent protein from eGFP/CFP etc pr 
                     mCherry etc"
     primer_bind     1321..1351
                     /label=NG-NIR1-IF
     primer_bind     1321..1351
                     /label=NG-NIR1-IF2
     primer_bind     1321..1335
                     /label=NG-NIR1-IF
     CDS             1339..1950
                     /codon_start=1
                     /label=Nir1
                     /translation="PANPREKWLRKRTQVKLRNVTANHRANDVIAAEDGPQVLVGRFMY
                     GPLDMVALTGEKVDILVMAEPSSGRWVHLDTEITNSSGRITYNVPRPRRLGVGVYPVKM
                     VVRGDQTCAMSYLTVLPRGMECVVFSIDGSFAASVSIMGSDPKVRPGAVDVVRHWQDLG
                     YMILYITGRPDMQKQRVVSWLSQHNFPQGMIFFSDGLVHDP"
     primer_bind     complement(1824..1845)
                     /label=Nir1-R784E-R
     primer_bind     1846..1870
                     /label=Nir1-R784E-F
     primer_bind     complement(1936..1950)
                     /label=Nir1-K820E-R
     CDS             1951..2193
                     /codon_start=1
                     /label=Nir1
                     /translation="LRQEAIFLRNLMQECFIKISAAYGSTKDISVYSVLGLPASQIFIV
                     GRPTKKYQTQCQFLSEGYAAHLAVLEASHRSRPKKN"
     primer_bind     1951..1971
                     /label=Nir1-K820E-F
     misc_feature    1960..1962
                     /label=K820E
     primer_bind     complement(2002..2019)
                     /label=Nir1-K843E-R
     primer_bind     2020..2047
                     /label=Nir1-K843E-F
     primer_bind     complement(2130..2145)
                     /label=Nir1-V885A-R
     primer_bind     2146..2166
                     /label=Nir1-V885A-F
     misc_feature    2155..2157
                     /label=A885V
                     /note="Mutation noted using the full-length protein 
                     numbering. Mutation is a natural varient with gnomAD ID: 
                     17-6455609-G-A"
     primer_bind     complement(2198..2222)
                     /label=NG-NIR1-IR2
     primer_bind     complement(2201..2212)
                     /label=NG-NIR1-IR
     primer_bind     2203..2231
                     /label=NG-NIR1-VF2
     misc_feature    2213..2300
                     /label=MCS
                     /note="MCS"
     primer_bind     2213..2232
                     /label=RepGFPf
     primer_bind     complement(2237..2257)
                     /label=INPPL1_V_R
     primer_bind     complement(2247..2266)
                     /label=EGFP_PIP5K1B_vR
     primer_bind     2259..2273
                     /label=INPPL1_V_F
     primer_bind     2271..2291
                     /label=EGFP_PIP5K1B_vF
     primer_bind     complement(2308..2327)
                     /label=SV40-pArev
     primer_bind     2361..2383
                     /label=A42
     misc_feature    2530..2985
                     /label=f1
                     /note="f1"
     promoter        3047..3075
                     /label=Bacterial
                     /note="Bacterial"
     promoter        3159..3388
                     /label=SV40early
                     /note="SV40early"
     misc_feature    3326..3461
                     /label=SV40
                     /note="SV40"
     misc_feature    3510..4304
                     /label=Kan/Neo
                     /note="Kan/Neo"
     misc_feature    4540..4558
                     /label=HSV-TK
                     /note="HSV-TK"
     misc_feature    4889..5532
                     /label=pUC
                     /note="pUC"
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 ccggtcgcca ccatggtgag caagggcgag gaggataaca tggcctctct cccagcgaca
      661 catgagttac acatctttgg ctccatcaac ggtgtggact ttgacatggt gggtcagggc
      721 accggcaatc caaatgatgg ttatgaggag ttaaacctga agtccaccaa gggtgacctc
      781 cagttctccc cctggattct ggtccctcat atcgggtatg gcttccatca gtacctgccc
      841 taccctgacg ggatgtcgcc tttccaggcc gccatggtag atggctccgg ataccaagtc
      901 catcgcacaa tgcagtttga agatggtgcc tcccttactg ttaactaccg ctacacctac
      961 gagggaagcc acatcaaagg agaggcccag gtgaagggga ctggtttccc tgctgacggt
     1021 cctgtgatga ccaactcgct gaccgctgcg gactggtgca ggtcgaagaa gacttacccc
     1081 aacgacaaaa ccatcatcag tacctttaag tggagttaca ccactggaaa tggcaagcgc
     1141 taccggagca ctgcgcggac cacctacacc tttgccaagc caatggcggc taactatctg
     1201 aagaaccagc cgatgtacgt gttccgtaag acggagctca agcactccaa gaccgagctc
     1261 aacttcaagg agtggcaaaa ggcctttacc gatgtgatgg gcatggacga gctgtacaag
     1321 ggaggctcag gaggcatgcc tgctaacccg cgtgaaaaat ggctgcgtaa acgtacccag
     1381 gttaaactgc gtaacgttac cgctaaccac cgtgcgaacg atgttatcgc ggcggaagat
     1441 ggtccgcagg ttctggttgg tcgtttcatg tacggtccgc tggacatggt tgctctgacc
     1501 ggtgaaaaag tggatatcct ggttatggca gaaccgtcta gcggtcgctg ggttcacctg
     1561 gacactgaaa tcaccaactc ctctggtcgt atcacctaca acgttccgcg tccgcgccgc
     1621 ctgggtgttg gtgtttaccc ggttaaaatg gttgttcgtg gtgaccagac ctgcgcgatg
     1681 tcttacctga ccgttctgcc gcgcggtatg gaatgtgttg tgttctctat cgacggttct
     1741 ttcgctgcga gcgtttctat catgggctct gacccgaaag ttcgtccggg tgcggttgac
     1801 gttgttcgtc actggcagga cctgggttac atgatcctgt acatcaccgg ccgtccggac
     1861 atgcagaaac agcgtgttgt ttcttggctg tctcagcaca acttcccgca gggtatgatc
     1921 ttcttctctg atggtctggt tcacgacccg ctgcgtcagg aagcgatctt cctgcgtaac
     1981 ctgatgcagg aatgcttcat caaaatctct gcggcgtacg gttctaccaa agacatctct
     2041 gtttactccg ttctgggtct gccggcttct cagatcttca tcgtgggtcg tccgaccaaa
     2101 aaataccaga cccagtgtca gttcctgtct gaaggttacg cggcgcacct ggctgttctg
     2161 gaagcttctc accgttctcg tccgaaaaag aactaagcgg ccgctcgagt cttccggact
     2221 cagatctcga gctcaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccac
     2281 cggatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     2341 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     2401 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2461 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2521 ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     2581 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     2641 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     2701 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     2761 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     2821 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     2881 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     2941 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     3001 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3061 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     3121 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3181 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3241 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3301 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3361 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3421 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     3481 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     3541 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3601 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3661 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     3721 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     3781 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     3841 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     3901 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     3961 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4021 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4081 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4141 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4201 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4261 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4321 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4381 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4441 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     4501 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     4561 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     4621 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     4681 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     4741 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     4801 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4861 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4921 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4981 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5041 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5101 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5161 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5221 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5281 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5341 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5401 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5461 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5521 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     5581 tgattctgtg gataaccgta ttaccgccat gcat
//
