LOCUS       sequence-123475-        5723 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5723)
  TITLE     de Lange Lab Vectors
REFERENCE   2  (bases 1 to 5723)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pLPC-N FLAG
FEATURES             Location/Qualifiers
     source          1..5723
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             120..589
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(841..864)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      864..873
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(904..923)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(905..923)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(968..1171)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(997..1017)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1172..1551
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1723..1742
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(1831..2430)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1914..1934)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2411..2430
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(2448..2864)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(2464..2480)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(2496..2518)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    3065..3264
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             3329..3798
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        3829..3933
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3934..4002,4003..4794)
                     /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"
     primer_bind     complement(4152..4171)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4954..5542
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5443..5462
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5696..5713
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaactatg
       61 gccatttaat gtaaatactt aagaaaaaaa accaaattaa ttttgataca tgctgcatgt
      121 gaaagacccc cgctgacggg tagtcaatca ctcagaggag accctcccaa ggaacagcga
      181 gaccacaagt cggatgcaac tgcaagaggg tttattggat acacgggtac ccgggcgact
      241 cagtcaatcg gaggactggc gccccgagtg aggggttgtg ggctctttta ttgagctcgg
      301 ggagcagaag cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc aaataaggca
      361 cagggtcatt tcaggtcctt ggggcaccct ggaaacatct gatggttctc tagaaactgc
      421 tgagggctgg accgcatctg gggaccatct gttcttggcc ctgagccggg gcaggaactg
      481 cttaccacag atatcctgtt tggcccatat tcagctgttc catctgttct tggccctgag
      541 ccggggcagg aactgcttac cacagatatc ctgtttggcc catattcagc tgttccatct
      601 gttcctgacc ttgatctgaa cttctctatt ctcagttatg tatttttcca tgccttgcaa
      661 aatggcgtta cttaagctag cttgccaaac ctacaggtgg ggtctttcat tccccccttt
      721 ttctggagac taaataaaat cttttatttt atcggccctc tagatgcatg ctcgagcggc
      781 cgccagtgtg atggatatct gcagaattcc agcacactgg cggccgttac tagtggatcc
      841 tttatcgtca tcgtctttgt agtccatggt ggctgatccg agctcggtac caagcttggg
      901 tctccctata gtgagtcgta ttaatttcga taagccagta agcagtgggt tctctagtta
      961 gccagagagc tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca
     1021 atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac
     1081 tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac
     1141 gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga
     1201 tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc
     1261 atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg
     1321 ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg
     1381 gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc
     1441 agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa
     1501 ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgt caacgcgtat
     1561 atctggcccg tacatcgcga agcagcgcaa aacgcctaac cctaagcaga ttcttcatgc
     1621 aattgtcggt caagccttgc cttgttgtag cttaaatttt gctcgcgcac tactcagcga
     1681 cctccaacac acaagcaggg agcagatact ggcttaacta tgcggcatca gagcagattg
     1741 tactgagagt gcaccatagg ggatcgggag atccgatgca tggggtcgtg cgctcctttc
     1801 ggtcgggcgc tgcgggtcgt ggggcgggcg tcaggcaccg ggcttgcggg tcatgcacca
     1861 ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta
     1921 gaaggggagg ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg cgcgctcggc
     1981 cgcctccact ccggggagca cgacggcgct gcccagaccc ttgccctggt ggtcgggcga
     2041 gacgccgacg gtggccagga accacgcggg ctccttgggc cggtgcggcg ccaggaggcc
     2101 ttccatctgt tgctgcgcgg ccagccggga accgctcaac tcggccatgc gcgggccgat
     2161 ctcggcgaac accgcccccg cttcgacgct ctccggcgtg gtccagaccg ccaccgcggc
     2221 gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga ggaagagttc
     2281 ttgcagctcg gtgacccgct cgatgtggcg gtccggatcg acggtgtggc gcgtggcggg
     2341 gtagtcggcg aacgcggcgg cgagggtgcg tacggccctg gggacgtcgt cgcgggtggc
     2401 gaggcgcacc gtgggcttgt actcggtcat ggtaagctga tccggccggc gcctagagaa
     2461 ggagtgaggg ctggataaag ggaggatcga ggcggggtcg aacgaggagg ttcaaggggg
     2521 agagacgggg cggatggagg aagaggaggc ggaggcttag ggtgtacaaa gggcttgacc
     2581 cagggagggg ggtcaaaagc caaggcttcc caggtcacga tgtaggggac ctggtctggg
     2641 tgtccatgcg ggccaggtga aaagaccttg atcttaacct gggtgatgag gtctcggtta
     2701 aaggtgccgt ctcgcggcca tccgacgtta aaggttggcc attctgcaga gcagaaggta
     2761 acccaacgtc tcttcttgac atctaccgac tggttgtgag cgatccgctc gacatctttc
     2821 cagtgaccta aggtcaaact taagggagtg gtaacagtct ggccctaatt ttcagacaaa
     2881 tacagaaaca cagtcagaca gagacaacac agaacgatgc tgcagcgctg cagcagacaa
     2941 gacgcgcggc ttcggttcca aaccgaaagc aaaaattcag acggaggcgg gaactgtttt
     3001 aggttctcgt ctcctaccag aaccacatat cctgacgggg tcggattcca catcgactcc
     3061 cttcctcagg tcgggccaca aaaacggccc ccaaagtccc tgggacgtct cccagggttg
     3121 cggccgggtg ttcagaactc gtcagttcca ccacgggtcc gccagataca gagctagtta
     3181 gctaactagt accgacgcag gcgcataaaa tcagtcatag acactagaca atcggacaga
     3241 cacagataag ttgctggcca gcttgcctcc cggtggtggg tcggtggtcc ctgggcaggg
     3301 gtctcccgat cccggacgag cccccaaatg aaagaccccc gctgacgggt agtcaatcac
     3361 tcagaggaga ccctcccaag gaacagcgag accacaagtc ggatgcaact gcaagagggt
     3421 ttattggata cacgggtacc cgggcgactc agtcaatcgg aggactggcg ccccgagtga
     3481 ggggttgtgg gctcttttat tgagctcggg gagcagaagc gcgcgaacag aagcgagaag
     3541 cgaactgatt ggttagttca aataaggcac agggtcattt caggtccttg gggcaccctg
     3601 gaaacatctg atggttctct agaaactgct gagggctgga ccgcatctgg ggaccatctg
     3661 ttcttggccc tgagccgggg caggaactgc ttaccacaga tatcctgttt ggcccatatt
     3721 cagctgttcc atctgttctt ggccctgagc cggggcagga actgcttacc acagatatcc
     3781 tgtttggccc atattcaggc tgcaggtggc acttttcggg gaaatgtgcg cggaacccct
     3841 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     3901 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     3961 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4021 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4081 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4141 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     4201 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4261 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     4321 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     4381 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     4441 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     4501 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     4561 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     4621 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     4681 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     4741 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     4801 gaccaagttt actcatatat actttagatt gatttgcggc cgcaaaagga tctaggtgaa
     4861 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4921 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4981 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5041 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5101 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5161 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5221 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5281 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5341 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5401 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5461 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5521 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5581 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5641 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     5701 gtcagtgagc gaggaagcgg aag
//
