pLYL122

LOCUS       pLYL122              8015 bp DNA     circular SYN 13-JAN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8015)
  AUTHORS   iGEM CPU-CHINA 2020
  TITLE     Direct Submission
  JOURNAL   Exported Jul 18, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..8015
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     235..251
                     /label=oMH18
     misc_feature    252..405
                     /label=Transcription blocker
                     /label=Transcription\blocker
     polyA_signal    252..300
                     /note="synthetic polyadenylation signal"
     misc_feature    314..405
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     misc_feature    439..525
                     /label=CRE reg_Regulatory_Seq
     misc_feature    450..457
                     /label=CRE
     misc_feature    486..493
                     /label=CRE*
     misc_feature    508..515
                     /label=CRE
     promoter        532..678
                     /label=P_TAL
     misc_recomb     694..727
                     /label=LoxP
     protein_bind    complement(694..727)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    787..2346
                     /label=SEAP
     CDS             787..2289
                     /codon_start=1
                     /product="secreted alkaline phosphatase from human 
                     placenta"
                     /label=SEAP
                     /note="The natural enzyme has a C-terminal transmembrane 
                     sequence that is replaced by a GPI anchor."
                     /translation="MLLLLLLLGLRLQLSLGIIPVEEENPDFWNREAAEALGAAKKLQP
                     AQTAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKTYNVDK
                     HVPDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGKSVGVV
                     TTTRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVILGGGRK
                     YMFRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLAKRQGARYVWNRTELMQASLDPSVTH
                     LMGLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRNPRGFFLFVEGGRIDHGHHESR
                     AYRALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSIFGLAPGKA
                     RDRKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGEDVAVFA
                     RGPQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTD"
     primer_bind     2278..2302
                     /label=183-WKG-599-F
     polyA_signal    2378..2602
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    2613..2734
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_recomb     2832..2865
                     /label=LoxP
     protein_bind    complement(2832..2865)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    2872..2920
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     misc_feature    2936..3004
                     /label=Secretory (IgK)
                     /label=nonstandard type:  signal
     CDS             3023..3535
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     misc_feature    3056..3058
                     /label=cyt-STOP (TGG->TGA)
     terminator      3542..3564
                     /label=SV40 late polyA
     polyA_signal    3612..3836
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3882..4310
                     /label=f1 bacteriophage origin of replication
                     /label=f1 bacteriophage origin of replication; arrow _f1 
                     ori
     primer_bind     complement(3896..3920)
                     /label=184-WKG-599-R
     promoter        4324..4653
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     misc_feature    4620..4625
                     /label=SDS
     CDS             4720..5514
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5688..5809
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5854..5874)
                     /label=M13-rev
     primer_bind     complement(5858..5874)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_binding    complement(5880..5902)
                     /label=LacO
     protein_bind    5882..5898
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5906..5936)
                     /label=lac promoter
     protein_bind    5951..5972
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    6078..7018
                     /label=ColE1
                     /note="ColE1"
     CDS             complement(7019..7879)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7880..7984)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgtggtactt
      241 ggagcggccg caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg
      301 aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa
      361 aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga taggtaccga
      421 gctcttacgc gtgctagcgc accagacagt gacgtcagct gccagatccc atggccgtca
      481 tactgtgacg tctttcagac accccattga cgtcaatggg agaacagatc tgccgccccg
      541 actgcatctg cgtgttcgaa ttcgccaatg acaagacgct gggcggggtt tgtgtcatca
      601 tagaactaaa gacatgcaaa tatatttctt ccggggacac cgccagcaaa cgcgagcaac
      661 gggccacggg gatgaagcag aaggctagcg cttataactt cgtatagcat acattatacg
      721 aagttatcgg cgcgccgcgg ccgcgatcaa ttcggtaccg aagttcctat tccgaattcg
      781 cccaccatgc tgctgctgct gctgctgctg ggcctgaggc tacagctctc cctgggcatc
      841 atcccagttg aggaggagaa cccggacttc tggaaccgcg aggcagccga ggccctgggt
      901 gccgccaaga agctgcagcc tgcacagaca gccgccaaga acctcatcat cttcctgggc
      961 gatgggatgg gggtgtctac ggtgacagct gccaggatcc taaaagggca gaagaaggac
     1021 aaactggggc ctgagatacc cctggccatg gaccgcttcc catatgtggc tctgtccaag
     1081 acatacaatg tagacaaaca tgtgccagac agtggagcca cagccacggc ctacctgtgc
     1141 ggggtcaagg gcaacttcca gaccattggc ttgagtgcag ccgcccgctt taaccagtgc
     1201 aacacgacac gcggcaacga ggtcatctcc gtgatgaatc gggccaagaa agcagggaag
     1261 tcagtgggag tggtaaccac cacacgagtg cagcacgcct cgccagccgg cacctacgcc
     1321 cacacggtga accgcaactg gtactcggac gccgacgtgc ctgcctcggc ccgccaggag
     1381 gggtgccagg acatcgctac gcagctcatc tccaacatgg acattgacgt gatcctaggt
     1441 ggaggccgaa agtacatgtt tcgcatggga accccagacc ctgagtaccc agatgactac
     1501 agccaaggtg ggaccaggct ggacgggaag aatctggtgc aggaatggct ggcgaagcgc
     1561 cagggtgccc ggtatgtgtg gaaccgcact gagctcatgc aggcttccct ggacccgtct
     1621 gtgacccatc tcatgggtct ctttgagcct ggagacatga aatacgagat ccaccgagac
     1681 tccacactgg acccctccct gatggagatg acagaggctg ccctgcgcct gctgagcagg
     1741 aacccccgcg gcttcttcct cttcgtggag ggtggtcgca tcgaccatgg tcatcatgaa
     1801 agcagggctt accgggcact gactgagacg atcatgttcg acgacgccat tgagagggcg
     1861 ggccagctca ccagcgagga ggacacgctg agcctcgtca ctgccgacca ctcccacgtc
     1921 ttctccttcg gaggctaccc cctgcgaggg agctccatct tcgggctggc ccctggcaag
     1981 gcccgggaca ggaaggccta cacggtcctc ctatacggaa acggtccagg ctatgtgctc
     2041 aaggacggcg cccggccgga tgttaccgag agcgagagcg ggagccccga gtatcggcag
     2101 cagtcagcag tgcccctgga cgaagagacc cacgcaggcg aggacgtggc ggtgttcgcg
     2161 cgcggcccgc aggcgcacct ggttcacggc gtgcaggagc agaccttcat agcgcacgtc
     2221 atggccttcg ccgcctgcct ggagccctac accgcctgcg acctggcgcc ccccgccggc
     2281 accaccgacg ccgcgcaccc gggttactct agagtcgggg cggccggccg cttcgagcag
     2341 acatgatcta gaggcctgca gggctgatca gcctcgactg tgccttctag ttgccagcca
     2401 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     2461 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     2521 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     2581 ggggatgcgg tgggctctat ggcttctgag gataagatac attgatgagt ttggacaaac
     2641 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2701 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2761 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc gcttggtacc gtctttccag
     2821 tggttaatta aataacttcg tatagcatac attatacgaa gttatggatc cgaacaaacg
     2881 acccaacacc cgtgcgtttt attctgtctt tttattgccg atcctcgagg ccaccatgac
     2941 tagtgagaca gacacactcc tgctatgggt actgctgctc tgggttccag gttccactgg
     3001 tgacgctagt ggtggttctg gtatggtctt cacactcgaa gatttcgttg gggactggcg
     3061 acagacagcc ggctacaacc tggaccaagt ccttgaacag ggaggtgtgt ccagtttgtt
     3121 tcagaatctc ggggtgtccg taactccgat ccaaaggatt gtcctgagcg gtgaaaatgg
     3181 gctgaagatc gacatccatg tcatcatccc gtatgaaggt ctgagcggcg accaaatggg
     3241 ccagatcgaa aaaattttta aggtggtgta ccctgtggat gatcatcact ttaaggtgat
     3301 cctgcactat ggcacactgg taatcgacgg ggttacgccg aacatgatcg actatttcgg
     3361 acggccgtat gaaggcatcg ccgtgttcga cggcaaaaag atcactgtaa cagggaccct
     3421 gtggaacggc aacaaaatta tcgacgagcg cctgatcaac cccgacggct ccctgctgtt
     3481 ccgagtaacc atcaacggag tgaccggctg gcggctgtgc gaacgcattc tggcgatcga
     3541 tagacatgat aagatacatt gatgtctaga agcttctaga gggcccgttt aaacccgctg
     3601 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3661 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3721 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3781 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     3841 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     3901 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3961 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4021 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4081 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4141 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4201 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4261 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     4321 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4381 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4441 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4501 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4561 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     4621 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     4681 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4741 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4801 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4861 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     4921 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4981 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     5041 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     5101 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     5161 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5221 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     5281 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5341 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5401 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5461 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5521 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5581 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5641 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     5701 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5761 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     5821 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5881 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5941 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     6001 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6061 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6121 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6181 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6241 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6301 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6361 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6421 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6481 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6541 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6601 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6661 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6721 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6781 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6841 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6901 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6961 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7021 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7081 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7141 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7201 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7261 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7321 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7381 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7441 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7501 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7561 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7621 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7681 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7741 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7801 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7861 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7921 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7981 cgcgcacatt tccccgaaaa gtgccacctg acgtc
//