LOCUS       pcDNA3.1(+)_EPB4        6602 bp    DNA     circular SYN 11-JUL-2025
DEFINITION  1146.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6602)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2025 from SnapGene 8.1.1
            https://www.snapgene.com
COMMENT     Ligation of : pcdna [5397  nt] : (NotI[978] / EcoRI[952]) to as1 
            [1197  nt] : (EcoRI[1] / NotI[1201])
FEATURES             Location/Qualifiers
     source          1..6602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    957..2153
                     /label=EPB41L4A-AS1
                     /note="note=""Fragment extracted from as1 [1197  nt] : 
                     (EcoRI[1] / NotI[1201])"
                     /note="color: #84a4c0; direction: RIGHT"
     polyA_signal    2202..2426
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      2472..2900
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        2914..3243
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             3310..4104
                     /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))"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4278..4399
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     promoter        complement(join(4496..4502,4503..4520,4521..4526))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 4496 .. 4502 / #ffffff / -10
                        2: 4503 .. 4520 / #ffffff
                        3: 4521 .. 4526 / #ffffff / -35"
     protein_bind    4541..4562
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(4850..5435)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(join(5606..6397,6398..6466))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 5606 .. 6397 / #ccffcc
                        2: 6398 .. 6466 / #ccffcc / signal sequence
                     Cleavage site after base 6397"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6467..6571)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
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 cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattctgt
      961 cgggtcccaa aatacgtggt ctaaagtcct ttgggctcct gccgctgcca gcactgatca
     1021 tatgctcttg accctggtcg ggtgggggga tgtctggata cctcctctgt ggaaagaact
     1081 ggccacacat ccgcagaggt gactcgggct gagaacccag gctcctcctc cctttggtga
     1141 cacgcccctc ggtccctcac tggcacttct ccctccggcc acacggcggc gtctcgccat
     1201 agcgcagcgg ccgatggtac agcccgctcc cccctcgcgc tctcggacag tgggtccttc
     1261 cacttgtaga aaagcattgt gggacggaag cctgtccttt cttccttttg gtgcgagctt
     1321 gctgtggttt ttgctctggg tcctctggga tggcgcctgg ctgtggccgc gtggtctctc
     1381 acgcaggggc gccgggcggg ggaacgcggc caccctgagt ctggtgagtc gactgcggcg
     1441 gcctgtgtcc gaagtgtccg gggccgtgaa caagggcagc ggcctggcct caggcctgcg
     1501 ttcccacgtt tggaaacggg gagcttcgtc gatttgtgtt tacatcatcg actatgccag
     1561 ggagttctcc agataagcct ggttttattt tcgtcagtga aaaggcctta ccgtataact
     1621 gactttatgc ttgccctgcc cccgtataaa ataacttaaa agcagcgtgc ctggttacag
     1681 ctgtttccac gtgcggtgct cgtcgggagt gatcacctac cctacaggtg gaagatggat
     1741 gcctgaagtg tagactgctg ctagctgaat accatctggg agcataaagg tgacctgaag
     1801 gatgtccttg gtgaggattt tgaaaatttg atcttcacaa gagttgcctg gatcatttga
     1861 aatttctggg agtctgagga gtactgacat aattacctgc tggagtctgt aaatacacat
     1921 ttaagacagt gaggatgtga ataaatatat taatgcactt tggcatttgt gttttaagtg
     1981 attaactgcc agaaacagct atttctaaaa agttataagg gaggaggggt tcttttttga
     2041 tcagtattca ctgctgtcac cataattaat agccttaaaa tagcttgtgt ttggcccaaa
     2101 gagaaatgta ctttcttcca gtgactcaaa aatcgtggct agaactagac tgcggccgct
     2161 cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     2221 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2281 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     2341 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     2401 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     2461 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2521 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     2581 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     2641 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     2701 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     2761 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     2821 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     2881 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     2941 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3001 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3061 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3121 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3181 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3241 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     3301 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3361 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3421 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3481 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3541 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3601 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3661 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3721 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     3781 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     3841 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     3901 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     3961 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4021 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4081 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4141 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4201 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4261 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4321 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4381 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     4441 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4501 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4561 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4621 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4681 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4741 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4801 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4861 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4921 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4981 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5041 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5101 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5161 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5221 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5281 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5341 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca
     5401 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5461 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5521 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5581 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5641 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5701 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5761 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5821 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5881 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5941 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6001 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6061 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6121 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6181 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6241 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6301 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6361 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6421 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6481 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6541 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     6601 tc
//
