LOCUS       Exported                5494 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 5494)
  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..5494
                     /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"
     misc_feature    1435..1441
                     /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
     CDS             1449..1469
                     /codon_start=1
                     /label=EGFP
                     /note="EGFP"
                     /translation="MVSKGEE"
     CDS             1449..1469
                     /codon_start=1
                     /label=EGFP
                     /note="EGFP"
                     /translation="MVSKGEE"
     misc_feature    1449..1469
                     /label=EGFP
                     /note="EGFP"
     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..2168)
                     /label=NG-insR
     primer_bind     complement(2137..2157)
                     /label=NG-insR
     CDS             2137..2156
                     /codon_start=1
                     /label=EGFP
                     /note="EGFP"
                     /translation="MDELYK"
     CDS             2137..2156
                     /codon_start=1
                     /label=EGFP
                     /note="EGFP"
                     /translation="MDELYK"
     misc_feature    2137..2156
                     /label=EGFP
                     /note="EGFP"
     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     complement(2188..2207)
                     /label=SV40pA-R
     misc_feature    2410..2865
                     /label=f1
                     /note="f1"
     misc_feature    2437..2865
                     /label=f1
                     /note="f1"
     misc_feature    complement(2452..2865)
                     /label=f1ori
                     /note="f1"
     misc_feature    2542..2848
                     /label=f1( )
                     /note="f1( )"
     promoter        2927..2955
                     /label=Bacterial
                     /note="Bacterial"
     promoter        3039..3268
                     /label=SV40early
                     /note="SV40early"
     misc_feature    3206..3341
                     /label=SV40
                     /note="SV40"
     misc_feature    3390..4184
                     /label=Kan/Neo
                     /note="Kan/Neo"
     misc_feature    4420..4438
                     /label=HSV-TK
                     /note="HSV-TK"
     misc_feature    4769..5412
                     /label=pUC
                     /note="pUC"
     primer_bind     complement(5475..5494)
                     /label=NGx-PHPLC-R
     primer_bind     join(5478..5494,1..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 tacaagtaaa
     2161 gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     2221 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     2281 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2341 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2401 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     2461 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     2521 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     2581 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     2641 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     2701 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     2761 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     2821 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     2881 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2941 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     3001 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3061 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3121 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3181 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3241 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3301 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     3361 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     3421 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3481 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3541 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     3601 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     3661 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     3721 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     3781 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     3841 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     3901 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     3961 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4021 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4081 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4141 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4201 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4261 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4321 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     4381 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     4441 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     4501 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     4561 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     4621 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     4681 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4741 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4801 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4861 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4921 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4981 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5041 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5101 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5161 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5221 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5281 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5341 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5401 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     5461 tgattctgtg gataaccgta ttaccgccat gcat
//
