LOCUS       Exported                5027 bp ds-DNA     circular SYN 13-APR-2024
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5027)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Dec 3, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5027
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     30..51
                     /label=1897a_SP6_pCS2
                     /note="color: black; sequence: GCTTGATTTAGGTGACACTATA"
     misc_feature    35..52
                     /label=Sp6_primer
                     /note="color: #a6acb3"
     primer_bind     64..79
                     /label=Primer 1
                     /note="color: black; sequence: CTTGTTCTTTTTGCAG"
     primer_bind     complement(80..101)
                     /label=HA MO
                     /note="color: black; sequence: TACGGATACATGGTGGCggatc"
     CDS             91..120
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="color: #cc99b2"
                     /translation="MYPYDVPDYA"
     primer_bind     91..111
                     /label=1057y
                     /note="color: black; sequence: 
                     CTGTTGTGGATGTATCCGTACGACGTACCG"
     CDS             121..837
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1: 121 .. 123 / #e1ff2f
                        2: 124 .. 126 / #e1ff2f / 1a
                        3: 127 .. 837 / #e1ff2f"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(121..145)
                     /label=1943_EYFP_MO
                     /note="color: black; sequence: ACAGCTCCTCGCCCTTGCTCACCAT"
     primer_bind     complement(621..630)
                     /label=HA MO
                     /note="color: black; sequence: TACGGATACATGGTGGCggatc"
     primer_bind     669..688
                     /label=1129
                     /note="color: black; sequence: CCAGCAGAACACCCCCATCG"
     primer_bind     810..830
                     /label=1103a
                     /note="color: black; sequence: CACTCTCGGCATGGACGAGCT"
     primer_bind     843..874
                     /label=Nluc_rec_fr
                     /note="color: black; sequence: 
                     taaGATCCCATCGATTCGCGAGCCTCTAGAACTATAGTGAGTCGTATTAC"
     misc_feature    856..874
                     /label=SnaBI homology arm
                     /note="color: #a6acb3"
     primer_bind     complement(856..873)
                     /label=recorder_T7_F
                     /note="color: black; sequence: 
                     TAATACGACTCACTATAGCGATCTgCCAacgcTattcTGGctG"
     misc_feature    875..907
                     /label=Patial Illumina TrueSeq adaptor
                     /note="color: #ffff00"
     primer_bind     875..907
                     /label=1985a_read1_se_oligo_illumina
                     /note="color: black; sequence: 
                     ACACTCTTTCCCTACACGACGCTCTTCCGATCT"
     primer_bind     875..907
                     /label=1694
                     /note="color: black; sequence: 
                     AATGATACGGCGACCACCGAGATCTACACCGCCATACACTCTTTCCCTACACGACGCTC
                     TTCCGATCT"
     primer_bind     895..907
                     /label=1985b_read2_seq_oligo_illumina
                     /note="color: black; sequence: 
                     GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT"
     misc_feature    896..926
                     /label=5p adaptor
                     /note="color: #ffff00"
     primer_bind     902..926
                     /label=recorder_T7_F
                     /note="color: black; sequence: 
                     TAATACGACTCACTATAGCGATCTgCCAacgcTattcTGGctG"
     primer_bind     902..926
                     /label=1842a_recor_lib_F
                     /note="color: black; sequence: CGATCTgCCAacgcTattcTGGctG"
     misc_feature    909..923
                     /label=Xcm1
                     /note="color: #a6acb3"
     misc_feature    931..960
                     /label=3p adaptor
                     /note="color: #ffff99"
     primer_bind     complement(931..955)
                     /label=1842b_lib_recorder_R
                     /note="color: black; sequence: GACgtgCCAgaatTgcgtTGGataG"
     misc_feature    935..949
                     /label=Xcm1
                     /note="color: #a6acb3"
     misc_feature    954..988
                     /label=recorder 5p
                     /note="color: #ff6600"
     misc_feature    989..993
                     /label=recorderloop
                     /note="color: #a6acb3"
     misc_feature    994..1028
                     /label=recorder 3p
                     /note="color: #00ccff"
     misc_feature    1029..1045
                     /label=NheI
                     /note="color: #a6acb3"
     misc_feature    1036..1069
                     /label=Partial Illumina TrueSeq adaptor
                     /note="color: #ffff99"
     primer_bind     complement(1036..1069)
                     /label=1985b_read2_seq_oligo_illumina
                     /note="color: black; sequence: 
                     GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT"
     primer_bind     complement(1036..1048)
                     /label=1985a_read1_se_oligo_illumina
                     /note="color: black; sequence: 
                     ACACTCTTTCCCTACACGACGCTCTTCCGATCT"
     primer_bind     complement(1036..1048)
                     /label=1694
                     /note="color: black; sequence: 
                     AATGATACGGCGACCACCGAGATCTACACCGCCATACACTCTTTCCCTACACGACGCTC
                     TTCCGATCT"
     primer_bind     1037..1069
                     /label=i7 sequencing primer
                     /note="color: black; sequence: 
                     GATCGGAAGAGCACACGTCTGAAcTCCAGTCAC"
     primer_bind     complement(1052..1069)
                     /label=1814
                     /note="color: black; sequence: 
                     CAAGCAGAAGACGGCATACGAGATcgtgatGTGACTGGAGTTCAGACG"
     primer_bind     complement(1057..1083)
                     /label=Primer 3
                     /note="color: black; sequence: 
                     TGCACGTCGAGCGGCCGGATCTACtacgtaGTGACTGGAgTTC"
     primer_bind     complement(1063..1095)
                     /label=Primer 2
                     /note="color: black; sequence: 
                     GCACGTCGAGCGGCCCTTATCATGTCTGGATCTACtacgtaGTGACTG"
     primer_bind     complement(1076..1112)
                     /label=rec_rev_nlc
                     /note="color: black; sequence: 
                     CAAACTCATCAATGTATCTTATCATGTCTGGATCTAC"
     misc_feature    1076..1095
                     /label=SnaBI homology arm
                     /note="color: #a6acb3"
     terminator      complement(1089..1208)
                     /label=SV40_PA_terminator
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     complement(1101..1122)
                     /label=1017b
                     /note="color: black; sequence: GTGGTTTGTCCAAACTCATCAA"
     misc_feature    complement(1103..1122)
                     /label=EBV_rev_primer
                     /note="color: #a6acb3; direction: LEFT"
     primer_bind     complement(1169..1208)
                     /label=1057d
                     /note="color: black; sequence: 
                     AGGAAGAAACGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGC"
     primer_bind     complement(1222..1255)
                     /label=Primer 4
                     /note="color: black; sequence: 
                     GCACGTCGAGCGGCCCCTGAACCTGAAACATAAAATGAATGCAATTG"
     primer_bind     complement(1258..1279)
                     /label=1897b_SV40_pCS2
                     /note="color: black; sequence: GAATTAAAAAACCTCCCACACC"
     primer_bind     complement(1267..1289)
                     /label=1373b
                     /note="color: black; sequence: CCGCGGCCGCGAATTAAAAAACC"
     promoter        complement(1325..1344)
                     /label=T3_promoter
                     /note="color: #ffffff; direction: LEFT"
     promoter        complement(1361..1379)
                     /label=M13_reverse_primer promoter
                     /label=M13_reverse_primer
                     /note="color: #ffffff; direction: LEFT"
     misc_feature    complement(1378..1400)
                     /label=M13_pUC_rev_primer
                     /note="color: #a6acb3; direction: LEFT"
     CDS             complement(2526..3386)
                     /codon_start=1
                     /label=AmpR
                     /label=ORF frame 1
                     /note="color: #993366"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(2526..3386)
                     /gene="Ampicillin"
                     /label=Ampicillin
                     /note="color: #a6acb3; direction: LEFT"
     promoter        complement(3428..3456)
                     /label=AmpR_promoter
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     3979..3992
                     /label=recorder_T7_F
                     /note="color: black; sequence: 
                     TAATACGACTCACTATAGCGATCTgCCAacgcTattcTGGctG"
     primer_bind     3979..3992
                     /label=1842a_recor_lib_F
                     /note="color: black; sequence: CGATCTgCCAacgcTattcTGGctG"
     misc_feature    4945..4986
                     /label=CMV_fwd_primer
                     /note="color: #a6acb3"
     primer_bind     join(5015..5027,1..9)
                     /label=1017a
                     /note="color: black; sequence: TCGTTTAGGGAACCGCCATTCT"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccgccacc atgtatccgt acgacgtacc ggattatgcg
      121 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
      181 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      241 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      301 ctcgtgacca ccttcggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag
      361 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      421 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      481 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      541 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
      601 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
      661 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      721 tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      781 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtag
      841 ctcgagcctc tagaactata gtgagtcgta ttacacactc tttccctaca cgacgctctt
      901 ccgatctgcc aacgctattc tggctgaatt ctatccaacg caattctggc acgtccaatc
      961 caatccaatc caatccaatc caatccaatt aaattagatt agattagatt agattagatt
     1021 agattagaca ggcctagatc ggaagagcac acgtctgaac tccagtcact acgtagtaga
     1081 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     1141 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     1201 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     1261 gtgggaggtt ttttaattcg cggccgcggc gccaatgcat tgggcccggt acccagcttt
     1321 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     1381 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccgggag cataaagtgt
     1441 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     1501 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     1561 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     1621 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     1681 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     1741 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     1801 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1861 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1921 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     1981 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2041 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2101 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2161 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     2221 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2281 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2341 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2401 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     2461 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     2521 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     2581 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     2641 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     2701 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     2761 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     2821 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     2881 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     2941 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3001 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     3061 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     3121 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     3181 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     3241 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     3301 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     3361 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     3421 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     3481 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
     3541 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     3601 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     3661 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     3721 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     3781 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     3841 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     3901 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3961 tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     4021 gggcctcttc gctattacgc cagtcgaccg ccaattcaat atggcgtata tggactcatg
     4081 ccaattcaat atggtggatc tggacctgtg ccaattcaat atggcgtata tggactcgtg
     4141 ccaattcaat atggtggatc tggaccccag ccaattcaat atggcggact tggcaccatg
     4201 ccaattcaat atggcggacc tggcactgtg ccaactgggg aggggtctac ttggcacggt
     4261 gccaagtttg aggaggggtc ttggccctgt gccaagtccg ccatattgaa ttggcatggt
     4321 gccaataatg gcggccatat tggctatatg ccaggatcaa tatataggca atatccaata
     4381 tggccctatg ccaatatggc tattggccag gttcaatact atgtattggc cctatgccat
     4441 atagtattcc atatatgggt tttcctattg acgtagatag cccctcccaa tgggcggtcc
     4501 catataccat atatggggct tcctaatacc gcccatagcc actcccccat tgacgtcaat
     4561 ggtctctata tatggtcttt cctattgacg tcatatgggc ggtcctattg acgtatatgg
     4621 cgcctccccc attgacgtca attacggtaa atggcccgcc tggctcaatg cccattgacg
     4681 tcaataggac cacccaccat tgacgtcaat gggatggctc attgcccatt catatccgtt
     4741 ctcacgcccc ctattgacgt caatgacggt aaatggccca cttggcagta catcaatatc
     4801 tattaatagt aacttggcaa gtacattact attggaagta cgccagggta cattggcagt
     4861 actcccattg acgtcaatgg cggtaaatgg cccgcgatgg ctgccaagta catccccatt
     4921 gacgtcaatg gggaggggca atgacgcaaa tgggcgttcc attgacgtaa atgggcggta
     4981 ggcgtgccta atgggaggtc tatataagca atgctcgttt agggaac
//
