pLZ560

LOCUS       pLZ560              6901 bp DNA     circular SYN 14-JAN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6901)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Jul 20, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6901
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    228
                     /label=E2F-1
     misc_feature    234..245
                     /label=VanO
                     /label=nonstandard type:  operator
     misc_feature    248..259
                     /label=VanO
                     /label=nonstandard type:  operator
     misc_feature    262..273
                     /label=VanO
                     /label=nonstandard type:  operator
     misc_feature    276..287
                     /label=VanO
                     /label=nonstandard type:  operator
     misc_feature    290..301
                     /label=VanO
                     /label=nonstandard type:  operator
     misc_feature    302..306
                     /label=E2F-1
     misc_feature    307..455
                     /label=hcmvmin
     misc_feature    377..391
                     /label=New Feature
     misc_feature    467..472
                     /label=NheI
     misc_feature    483..487
                     /label=New Feature
     misc_feature    483..487
                     /label=New Feature
     CDS             498..2000
                     /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"
     polyA_signal    2049..2057
                     /label=SV40 late polyA
     sig_peptide     2055..2114
                     /label=P2A
     CDS             2058..2114
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2115..2444
                     /label=mINS
     polyA_signal    2498..2722
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2768..3196
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     3067..3086
                     /label=189-WKG-612-F
     promoter        3210..3539
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3606..4400
                     /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"
     primer_bind     complement(3650..3677)
                     /label=188-WKG-610-R
     polyA_signal    4574..4695
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4740..4760)
                     /label=M13-rev
     primer_bind     complement(4744..4760)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_binding    complement(4766..4788)
                     /label=LacO
     protein_bind    4768..4784
                     /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(4792..4822)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        complement(4793..4822)
                     /label=lac
     protein_bind    4837..4858
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    4964..5904
                     /label=ColE1
                     /note="ColE1"
     rep_origin      complement(5146..5734)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5905..6765)
                     /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(6766..6870)
                     /gene="bla"
                     /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 cgtattggat
      241 ccaatgcatt ggatccaatg gattggatcc aatcgattgg atccaattga ttggatccaa
      301 tctagctcga gctcggtacc cgggtcgagg taggcgtgta cggtgggagg cctatataag
      361 cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
      421 ccatagaaga caccgggacc gatccagcct ccgcggcccc gaattcgcta gcgtttaaac
      481 ttaagcttag cgccaccatg ctgctgctgc tgctgctgct gggcctgagg ctacagctct
      541 ccctgggcat catcccagtt gaggaggaga acccggactt ctggaaccgc gaggcagccg
      601 aggccctggg tgccgccaag aagctgcagc ctgcacagac agccgccaag aacctcatca
      661 tcttcctggg cgatgggatg ggggtgtcta cggtgacagc tgccaggatc ctaaaagggc
      721 agaagaagga caaactgggg cctgagatac ccctggccat ggaccgcttc ccatatgtgg
      781 ctctgtccaa gacatacaat gtagacaaac atgtgccaga cagtggagcc acagccacgg
      841 cctacctgtg cggggtcaag ggcaacttcc agaccattgg cttgagtgca gccgcccgct
      901 ttaaccagtg caacacgaca cgcggcaacg aggtcatctc cgtgatgaat cgggccaaga
      961 aagcagggaa gtcagtggga gtggtaacca ccacacgagt gcagcacgcc tcgccagccg
     1021 gcacctacgc ccacacggtg aaccgcaact ggtactcgga cgccgacgtg cctgcctcgg
     1081 cccgccagga ggggtgccag gacatcgcta cgcagctcat ctccaacatg gacattgacg
     1141 tgatcctagg tggaggccga aagtacatgt ttcgcatggg aaccccagac cctgagtacc
     1201 cagatgacta cagccaaggt gggaccaggc tggacgggaa gaatctggtg caggaatggc
     1261 tggcgaagcg ccagggtgcc cggtatgtgt ggaaccgcac tgagctcatg caggcttccc
     1321 tggacccgtc tgtgacccat ctcatgggtc tctttgagcc tggagacatg aaatacgaga
     1381 tccaccgaga ctccacactg gacccctccc tgatggagat gacagaggct gccctgcgcc
     1441 tgctgagcag gaacccccgc ggcttcttcc tcttcgtgga gggtggtcgc atcgaccatg
     1501 gtcatcatga aagcagggct taccgggcac tgactgagac gatcatgttc gacgacgcca
     1561 ttgagagggc gggccagctc accagcgagg aggacacgct gagcctcgtc actgccgacc
     1621 actcccacgt cttctccttc ggaggctacc ccctgcgagg gagctccatc ttcgggctgg
     1681 cccctggcaa ggcccgggac aggaaggcct acacggtcct cctatacgga aacggtccag
     1741 gctatgtgct caaggacggc gcccggccgg atgttaccga gagcgagagc gggagccccg
     1801 agtatcggca gcagtcagca gtgcccctgg acgaagagac ccacgcaggc gaggacgtgg
     1861 cggtgttcgc gcgcggcccg caggcgcacc tggttcacgg cgtgcaggag cagaccttca
     1921 tagcgcacgt catggccttc gccgcctgcc tggagcccta caccgcctgc gacctggcgc
     1981 cccccgccgg caccaccgac gccgcgcacc cgggttactc tagagtcggg gcggccggcc
     2041 gcttcgagca gacaggagca accaactttt ccctgctgaa gcaggcaggc gacgtggagg
     2101 agaatcctgg acccatggcc ctgtggatgc gcttcctgcc cctgctggcc ctgctcgtcc
     2161 tctgggagcc caagcctgcc caggcttttg tcaaacagca cctttgtggt cctcacctgg
     2221 tggaggctct gtacctggtg tgtggggaac gtggtttctt ctacacaccc aagtcccgtc
     2281 gtaaaaggga ggacccgcaa gtgccacaac tggagctggg tggaggcccg gaggccgggg
     2341 atcttcagac cttggcactg gaggttgccc ggcagaagcg tggcattgtg gatcagtgct
     2401 gcaccagcat ctgctccctc taccaactgg agaactactg caactgagcg gccgctcgag
     2461 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     2521 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     2581 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     2641 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     2701 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     2761 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2821 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     2881 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     2941 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3001 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3061 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3121 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3181 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3241 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3301 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3361 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3421 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     3481 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     3541 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     3601 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3661 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3721 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     3781 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     3841 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3901 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     3961 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4021 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4081 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     4141 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4201 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4261 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4321 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4381 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     4441 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4501 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4561 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4621 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4681 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     4741 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4801 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4861 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4921 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4981 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5041 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5101 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5161 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5221 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5281 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5341 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5401 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5461 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5521 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5581 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5641 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5701 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5761 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5821 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5881 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5941 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6001 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6061 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6121 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6181 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6241 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6301 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6361 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6421 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6481 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6541 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6601 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6661 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6721 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6781 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6841 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6901 c
//