LOCUS       Exported                6217 bp ds-DNA     circular SYN 10-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6217)
  AUTHORS   Jonathan Pacheco
  TITLE     Direct Submission
  JOURNAL   Exported Friday, May 19, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6217
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..155
                     /label=CMV
                     /note="CMV"
     misc_feature    1..155
                     /label=CMVd
     misc_feature    204..713
                     /label=PH-PLCD1
                     /note="PLCD_GFP"
     primer_bind     complement(693..740)
                     /label=PHPLC-NGxR
     primer_bind     complement(725..751)
                     /label=NG-insR
     primer_bind     725..748
                     /label=NG-insF
     CDS             726..1433
                     /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"
     misc_feature    726..1433
                     /label=mNG
     misc_feature    726..746
                     /label=EGFP
                     /note="EGFP"
     CDS             726..746
                     /codon_start=1
                     /label=EGFP
                     /note="EGFP"
                     /translation="MVSKGEE"
     primer_bind     726..743
                     /label=FPi-F
                     /note="Amplify fluorescent protein from eGFP/CFP etc pr 
                     mCherry etc"
     primer_bind     726..743
                     /label=NGx-PHPLC-F
     primer_bind     729..746
                     /label=FPi-F2.0
     primer_bind     complement(729..746)
                     /label=FPv-R
                     /note="Amplify vector without eGFP/CFP etc pr mCherry etc. 
                     Use to exchange with FPi primers."
     regulatory      981..990
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     1315..1335
                     /label=NeonGFWD1
     primer_bind     complement(1414..1474)
                     /label=NG-insR
     misc_feature    1414..1433
                     /label=EGFP
                     /note="EGFP"
     CDS             1414..1433
                     /codon_start=1
                     /label=EGFP
                     /note="EGFP"
                     /translation="MDELYK"
     primer_bind     complement(1414..1433)
                     /label=FPi-R
                     /note="Amplify fluorescent protein from eGFP/CFP etc pr 
                     mCherry etc"
     primer_bind     1414..1433
                     /label=FPv-F
                     /note="Amplify vector without eGFP/CFP etc pr mCherry etc. 
                     Use to exchange with FPi primers."
     primer_bind     1431..1457
                     /label=EGFP-F
     misc_feature    1434..1435
                     /label=MCS
                     /note="MCS"
     primer_bind     1440..1457
                     /label=EGFP-F
     primer_bind     1447..1471
                     /label=NG-insF
     primer_bind     complement(1448..1463)
                     /label=PHPLC-NGxR
     CDS             1449..2156
                     /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"
     misc_feature    1449..2156
                     /label=mNG
     misc_feature    1449..1469
                     /label=EGFP
                     /note="EGFP"
     CDS             1449..1469
                     /codon_start=1
                     /label=EGFP
                     /note="EGFP"
                     /translation="MVSKGEE"
     primer_bind     1449..1466
                     /label=FPi-F
                     /note="Amplify fluorescent protein from eGFP/CFP etc pr 
                     mCherry etc"
     primer_bind     1449..1466
                     /label=NGx-PHPLC-F
     primer_bind     1452..1469
                     /label=FPi-F2.0
     primer_bind     complement(1452..1469)
                     /label=FPv-R
                     /note="Amplify vector without eGFP/CFP etc pr mCherry etc. 
                     Use to exchange with FPi primers."
     regulatory      1704..1713
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     2038..2058
                     /label=NeonGFWD1
     primer_bind     complement(2137..2197)
                     /label=NG-insR
     misc_feature    2137..2156
                     /label=EGFP
                     /note="EGFP"
     CDS             2137..2156
                     /codon_start=1
                     /label=EGFP
                     /note="EGFP"
                     /translation="MDELYK"
     primer_bind     complement(2137..2156)
                     /label=FPi-R
                     /note="Amplify fluorescent protein from eGFP/CFP etc pr 
                     mCherry etc"
     primer_bind     2137..2156
                     /label=FPv-F
                     /note="Amplify vector without eGFP/CFP etc pr mCherry etc. 
                     Use to exchange with FPi primers."
     primer_bind     2154..2180
                     /label=EGFP-F
     misc_feature    2157..2158
                     /label=MCS
                     /note="MCS"
     misc_feature    2158..2164
                     /label=MCS
                     /note="MCS"
     primer_bind     2163..2180
                     /label=EGFP-F
     primer_bind     2170..2194
                     /label=NG-insF
     primer_bind     complement(2171..2186)
                     /label=PHPLC-NGxR
     CDS             2172..2879
                     /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"
     misc_feature    2172..2879
                     /label=mNG
     CDS             2172..2192
                     /codon_start=1
                     /label=EGFP
                     /note="EGFP"
                     /translation="MVSKGEE"
     CDS             2172..2192
                     /codon_start=1
                     /label=EGFP
                     /note="EGFP"
                     /translation="MVSKGEE"
     misc_feature    2172..2192
                     /label=EGFP
                     /note="EGFP"
     primer_bind     2172..2189
                     /label=FPi-F
                     /note="Amplify fluorescent protein from eGFP/CFP etc pr 
                     mCherry etc"
     primer_bind     2172..2189
                     /label=NGx-PHPLC-F
     primer_bind     2175..2192
                     /label=FPi-F2.0
     primer_bind     complement(2175..2192)
                     /label=FPv-R
                     /note="Amplify vector without eGFP/CFP etc pr mCherry etc. 
                     Use to exchange with FPi primers."
     regulatory      2427..2436
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     2761..2781
                     /label=NeonGFWD1
     primer_bind     complement(2860..2891)
                     /label=NG-insR
     primer_bind     complement(2860..2880)
                     /label=NG-insR
     CDS             2860..2879
                     /codon_start=1
                     /label=EGFP
                     /note="EGFP"
                     /translation="MDELYK"
     CDS             2860..2879
                     /codon_start=1
                     /label=EGFP
                     /note="EGFP"
                     /translation="MDELYK"
     misc_feature    2860..2879
                     /label=EGFP
                     /note="EGFP"
     primer_bind     complement(2860..2879)
                     /label=FPi-R
                     /note="Amplify fluorescent protein from eGFP/CFP etc pr 
                     mCherry etc"
     primer_bind     2860..2879
                     /label=FPv-F
                     /note="Amplify vector without eGFP/CFP etc pr mCherry etc. 
                     Use to exchange with FPi primers."
     primer_bind     complement(2911..2930)
                     /label=SV40pA-R
     misc_feature    3133..3588
                     /label=f1
                     /note="f1"
     misc_feature    3160..3588
                     /label=f1
                     /note="f1"
     misc_feature    complement(3175..3588)
                     /label=f1ori
                     /note="f1"
     misc_feature    3265..3571
                     /label=f1( )
                     /note="f1( )"
     promoter        3650..3678
                     /label=Bacterial
                     /note="Bacterial"
     promoter        3762..3991
                     /label=SV40early
                     /note="SV40early"
     misc_feature    3929..4064
                     /label=SV40
                     /note="SV40"
     misc_feature    4113..4907
                     /label=Kan/Neo
                     /note="Kan/Neo"
     misc_feature    5143..5161
                     /label=HSV-TK
                     /note="HSV-TK"
     misc_feature    5492..6135
                     /label=pUC
                     /note="pUC"
     primer_bind     complement(6198..6217)
                     /label=NGx-PHPLC-R
     primer_bind     join(6201..6217,1..21)
                     /label=PHPLC-NGxF
ORIGIN
        1 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
       61 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      121 atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg actcagatct
      181 cgagctcaag cttcgaattc ggcatggact cgggccggga cttcctgacc ctgcacggcc
      241 tacaggatga tgaggatcta caggcgctgc tgaagggcag ccagctcctg aaggtgaagt
      301 ccagctcatg gaggagagag cggttctaca agttgcagga ggactgcaag accatctggc
      361 aggagtcccg caaggtcatg cggaccccgg agtcccagct gttctccatc gaggacattc
      421 aggaggtgcg aatggggcac cgcacggagg gtctggagaa gttcgcccgt gatgtgcccg
      481 aggaccgctg cttctccatt gtcttcaagg accagcgcaa tacactagac ctcatcgccc
      541 catcgccagc tgatgcccag cactgggtgc tggggctgca caagatcatc caccactcag
      601 gctccatgga ccagcgtcag aagctacagc actggattca ctcctgcttg cgaaaagctg
      661 acaaaaacaa ggacaacaag atgagcttca aggagctgca gaacttcctg aagggggtgg
      721 gtggcatggt gagcaagggc gaggaggata acatggcctc tctcccagcg acacatgagt
      781 tacacatctt tggctccatc aacggtgtgg actttgacat ggtgggtcag ggcaccggca
      841 atccaaatga tggttatgag gagttaaacc tgaagtccac caagggtgac ctccagttct
      901 ccccctggat tctggtccct catatcgggt atggcttcca tcagtacctg ccctaccctg
      961 acgggatgtc gcctttccag gccgccatgg tagatggctc cggataccaa gtccatcgca
     1021 caatgcagtt tgaagatggt gcctccctta ctgttaacta ccgctacacc tacgagggaa
     1081 gccacatcaa aggagaggcc caggtgaagg ggactggttt ccctgctgac ggtcctgtga
     1141 tgaccaactc gctgaccgct gcggactggt gcaggtcgaa gaagacttac cccaacgaca
     1201 aaaccatcat cagtaccttt aagtggagtt acaccactgg aaatggcaag cgctaccgga
     1261 gcactgcgcg gaccacctac acctttgcca agccaatggc ggctaactat ctgaagaacc
     1321 agccgatgta cgtgttccgt aagacggagc tcaagcactc caagaccgag ctcaacttca
     1381 aggagtggca aaaggccttt accgatgtga tgggcatgga cgagctgtac aagtcaccgg
     1441 tcgccaccat ggtgagcaag ggcgaggagg ataacatggc ctctctccca gcgacacatg
     1501 agttacacat ctttggctcc atcaacggtg tggactttga catggtgggt cagggcaccg
     1561 gcaatccaaa tgatggttat gaggagttaa acctgaagtc caccaagggt gacctccagt
     1621 tctccccctg gattctggtc cctcatatcg ggtatggctt ccatcagtac ctgccctacc
     1681 ctgacgggat gtcgcctttc caggccgcca tggtagatgg ctccggatac caagtccatc
     1741 gcacaatgca gtttgaagat ggtgcctccc ttactgttaa ctaccgctac acctacgagg
     1801 gaagccacat caaaggagag gcccaggtga aggggactgg tttccctgct gacggtcctg
     1861 tgatgaccaa ctcgctgacc gctgcggact ggtgcaggtc gaagaagact taccccaacg
     1921 acaaaaccat catcagtacc tttaagtgga gttacaccac tggaaatggc aagcgctacc
     1981 ggagcactgc gcggaccacc tacacctttg ccaagccaat ggcggctaac tatctgaaga
     2041 accagccgat gtacgtgttc cgtaagacgg agctcaagca ctccaagacc gagctcaact
     2101 tcaaggagtg gcaaaaggcc tttaccgatg tgatgggcat ggacgagctg tacaagtcac
     2161 cggtcgccac catggtgagc aagggcgagg aggataacat ggcctctctc ccagcgacac
     2221 atgagttaca catctttggc tccatcaacg gtgtggactt tgacatggtg ggtcagggca
     2281 ccggcaatcc aaatgatggt tatgaggagt taaacctgaa gtccaccaag ggtgacctcc
     2341 agttctcccc ctggattctg gtccctcata tcgggtatgg cttccatcag tacctgccct
     2401 accctgacgg gatgtcgcct ttccaggccg ccatggtaga tggctccgga taccaagtcc
     2461 atcgcacaat gcagtttgaa gatggtgcct cccttactgt taactaccgc tacacctacg
     2521 agggaagcca catcaaagga gaggcccagg tgaaggggac tggtttccct gctgacggtc
     2581 ctgtgatgac caactcgctg accgctgcgg actggtgcag gtcgaagaag acttacccca
     2641 acgacaaaac catcatcagt acctttaagt ggagttacac cactggaaat ggcaagcgct
     2701 accggagcac tgcgcggacc acctacacct ttgccaagcc aatggcggct aactatctga
     2761 agaaccagcc gatgtacgtg ttccgtaaga cggagctcaa gcactccaag accgagctca
     2821 acttcaagga gtggcaaaag gcctttaccg atgtgatggg catggacgag ctgtacaagt
     2881 aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     2941 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     3001 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3061 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3121 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     3181 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     3241 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     3301 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     3361 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     3421 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     3481 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     3541 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     3601 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3661 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     3721 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3781 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3841 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3901 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3961 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     4021 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     4081 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4141 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4201 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4261 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     4321 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4381 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4441 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4501 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4561 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4621 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4681 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4741 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4801 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4861 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     4921 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4981 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5041 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     5101 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     5161 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     5221 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     5281 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     5341 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     5401 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5461 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5521 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5581 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5641 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     5701 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5761 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5821 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5881 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5941 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6001 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6061 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6121 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6181 ccctgattct gtggataacc gtattaccgc catgcat
//
