LOCUS       Exported                5113 bp ds-DNA     circular SYN 20-JUL-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5113)
  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..5113
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    35..52
                     /label=Sp6_primer
                     /note="color: #a6acb3"
     primer_bind     91..111
                     /label=1057y
                     /note="color: black; sequence: 
                     CTGTTGTGGATGTATCCGTACGACGTACCG"
     CDS             94..120
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="color: #cc99b2"
                     /translation="YPYDVPDYA"
     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     669..688
                     /label=1129
                     /note="color: black; sequence: CCAGCAGAACACCCCCATCG"
     primer_bind     810..830
                     /label=1103a
                     /note="color: black; sequence: CACTCTCGGCATGGACGAGCT"
     primer_bind     856..895
                     /label=1745a_3xBoxB_SnaB1
                     /note="color: black; sequence: 
                     CTATAGTGAGTCGTATTACACACTCTTTCCCTACACGACG"
     misc_feature    856..874
                     /label=SnaBI homology arm
                     /note="color: #a6acb3"
     misc_feature    875..907
                     /label=Patial Illumina TrueSeq adaptor
                     /note="color: #ffff00"
     primer_bind     875..907
                     /label=1694
                     /note="color: black; sequence: 
                     AATGATACGGCGACCACCGAGATCTACACCGCCATACACTCTTTCCCTACACGACGCTC
                     TTCCGATCT"
     primer_bind     complement(909..927)
                     /label=1745b_3xBoxB_SnaB1
                     /note="color: black; sequence: 
                     CTTATCATGTCTGGATCTACcagCCAgaatTgcgtTGG"
     misc_feature    909..923
                     /label=Xcm1
                     /note="color: #a6acb3"
     protein_bind    927..945
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
                     /note="color: #31849b"
     primer_bind     complement(931..1031)
                     /label=BoxB_reverse
                     /note="color: black; sequence: 
                     CAgaatTgcgtTGGatagggcccttcttcagggccctatatgggcccttcttcagggcc
                     cagtttagtagttggacttagtcgaggggcccttcttcagg"
     protein_bind    971..989
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
                     /note="color: #31849b"
     protein_bind    995..1013
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
                     /note="color: #31849b"
     primer_bind     complement(1018..1033)
                     /label=1745b_3xBoxB_SnaB1
                     /note="color: black; sequence: 
                     CTTATCATGTCTGGATCTACcagCCAgaatTgcgtTGG"
     misc_feature    1018..1032
                     /label=Xcm1
                     /note="color: #a6acb3"
     misc_feature    1037..1071
                     /label=recorder 5p
                     /note="color: #ff6600"
     misc_feature    1072..1076
                     /label=recorderloop
                     /note="color: #a6acb3"
     misc_feature    1077..1111
                     /label=recorder 3p
                     /note="color: #00ccff"
     misc_feature    1112..1128
                     /label=NheI
                     /note="color: #a6acb3"
     misc_feature    1119..1152
                     /label=Partial Illumina TrueSeq adaptor
                     /note="color: #ffff99"
     primer_bind     complement(1128..1150)
                     /label=1744_illumina_rev
                     /note="color: black; sequence: GACTGGACTTCAGACGTGTGCTC"
     misc_feature    1159..1178
                     /label=SnaBI homology arm
                     /note="color: #a6acb3"
     primer_bind     complement(1159..1178)
                     /label=1745b_3xBoxB_SnaB1
                     /note="color: black; sequence: 
                     CTTATCATGTCTGGATCTACcagCCAgaatTgcgtTGG"
     terminator      complement(1172..1291)
                     /label=SV40_PA_terminator
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     complement(1184..1205)
                     /label=1017b
                     /note="color: black; sequence: GTGGTTTGTCCAAACTCATCAA"
     misc_feature    complement(1186..1205)
                     /label=EBV_rev_primer
                     /note="color: #a6acb3; direction: LEFT"
     primer_bind     complement(1252..1291)
                     /label=1057d
                     /note="color: black; sequence: 
                     AGGAAGAAACGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGC"
     primer_bind     complement(1350..1372)
                     /label=1373b
                     /note="color: black; sequence: CCGCGGCCGCGAATTAAAAAACC"
     promoter        complement(1408..1427)
                     /label=T3_promoter
                     /note="color: #ffffff; direction: LEFT"
     promoter        complement(1444..1462)
                     /label=M13_reverse_primer promoter
                     /label=M13_reverse_primer
                     /note="color: #ffffff; direction: LEFT"
     misc_feature    complement(1461..1483)
                     /label=M13_pUC_rev_primer
                     /note="color: #a6acb3; direction: LEFT"
     CDS             complement(2609..3469)
                     /codon_start=1
                     /label=AmpR
                     /label=ORF frame 1
                     /note="color: #993366"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(2609..3469)
                     /gene="Ampicillin"
                     /label=Ampicillin
                     /note="color: #a6acb3; direction: LEFT"
     promoter        complement(3511..3539)
                     /label=AmpR_promoter
                     /note="color: #ffffff; direction: LEFT"
     misc_feature    5031..5072
                     /label=CMV_fwd_primer
                     /note="color: #a6acb3"
     primer_bind     join(5101..5113,1..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 tggctggggc cctgaagaag ggcccctcga ctaagtccaa
      961 ctactaaact gggccctgaa gaagggccca tatagggccc tgaagaaggg cccctatcca
     1021 acgcaattct ggcacgtcca atccaatcca atccaatcca atccaatcca attaaattag
     1081 attagattag attagattag attagattag acaggccttg atcggaagag cacacgtctg
     1141 aagtccagtc actacgtagt agatccagac atgataagat acattgatga gtttggacaa
     1201 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     1261 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     1321 atgtttcagg ttcaggggga ggtgtgggag gttttttaat tcgcggccgc ggcgccaatg
     1381 cattgggccc ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
     1441 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     1501 tacgagccgg gagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     1561 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     1621 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     1681 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     1741 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     1801 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1861 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1921 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1981 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2041 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2101 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2161 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2221 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2281 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2341 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2401 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2461 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2521 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2581 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     2641 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2701 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     2761 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     2821 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     2881 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     2941 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3001 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3061 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3121 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3181 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3241 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3301 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3361 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3421 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3481 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3541 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     3601 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     3661 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     3721 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     3781 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     3841 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     3901 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     3961 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     4021 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa
     4081 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagtcga ccatagccaa
     4141 ttcaatatgg cgtatatgga ctcatgccaa ttcaatatgg tggatctgga cctgtgccaa
     4201 ttcaatatgg cgtatatgga ctcgtgccaa ttcaatatgg tggatctgga ccccagccaa
     4261 ttcaatatgg cggacttggc accatgccaa ttcaatatgg cggacttggc actgtgccaa
     4321 ctggggaggg gtctacttgg cacggtgcca agtttgagga ggggtcttgg ccctgtgcca
     4381 agtccgccat attgaattgg catggtgcca ataatggcgg ccatattggc tatatgccag
     4441 gatcaatata taggcaatat ccaatatggc cctatgccaa tatggctatt ggccaggttc
     4501 aatactatgt attggcccta tgccatatag tattccatat atgggttttc ctattgacgt
     4561 agatagcccc tcccaatggg cggtcccata taccatatat ggggcttcct aataccgccc
     4621 atagccactc ccccattgac gtcaatggtc tctatatatg gtctttccta ttgacgtcat
     4681 atgggcggtc ctattgacgt atatggcgcc tcccccattg acgtcaatta cggtaaatgg
     4741 cccgcctggc tcaatgccca ttgacgtcaa taggaccacc caccattgac gtcaatggga
     4801 tggctcattg cccattcata tccgttctca cgccccctat tgacgtcaat gacggtaaat
     4861 ggcccacttg gcagtacatc aatatctatt aatagtaact tggcaagtac attactattg
     4921 gaaggacgcc agggtacatt ggcagtactc ccattgacgt caatggcggt aaatggcccg
     4981 cgatggctgc caagtacatc cccattgacg tcaatgggga ggggcaatga cgcaaatggg
     5041 cgttccattg acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata taagcaatgc
     5101 tcgtttaggg aac
//
