LOCUS       sequence-260470-        6034 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 6034)
  TITLE     pLAI-Delta-mls
REFERENCE   2  (bases 1 to 6034)
  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: CMV-SEAP
FEATURES             Location/Qualifiers
     source          1..6034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(35..1552)
                     /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="MLGPCMLLLLLLLGLRLQLSLGIIPVEEENPDFWNREAAEALGAA
                     KKLQPAQTAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKT
                     YNVDKHVPDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGK
                     SVGVVTTTRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVIL
                     GGGRKYMFRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLAKRQGARYVWNRTELMQASLD
                     PSVTHLMGLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRNPRGFFLFVEGGRIDHG
                     HHESRAYRALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSIFGL
                     APGKARDRKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGED
                     VAVFARGPQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTD"
     promoter        complement(1717..1920)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1746..1766)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1921..2300
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(2533..2552)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2534..2552)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2559..2576)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2559..2575)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2568..2590)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2717..3169
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2801..2820)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3011..3032
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3195..3299
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3300..3368,3369..4160)
                     /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(3518..3537)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4331..4919
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4820..4839
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5073..5090
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5207..5228
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5243..5248,5249..5266,5267..5273)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5281..5297
                     /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)."
     primer_bind     5286..5308
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5305..5321
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5305..5321
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5342..5360
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    5406..5540
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5430..5449)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5484..5503
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 acgcggggac cacgggttaa cccgggtgcg cggcgtcggt ggtgccggcg gggggcgcca
       61 ggtcgcaggc ggtgtagggc tccaggcagg cggcgaaggc catgacgtgc gctatgaagg
      121 tctgctcctg cacgccgtga accaggtgcg cctgcgggcc gcgcgcgaac accgccacgt
      181 cctcgcctgc gtgggtctct tcgtccaggg gcactgctga ctgctgccga tactcggggc
      241 tcccgctctc gctctcggta acatccggcc gggcgccgtc cttgagcaca tagcctggac
      301 cgtttccgta taggaggacc gtgtaggcct tcctgtcccg ggccttgcca ggggccagcc
      361 cgaagatgga gctccctcgc agggggtagc ctccgaagga gaagacgtgg gagtggtcgg
      421 cagtgacgag gctcagcgtg tcctcctcgc tggtgagctg gcccgccctc tcaatggcgt
      481 cgtcgaacat gatcgtctca gtcagtgccc ggtaagccct gctttcatga tgaccatggt
      541 cgatgcgacc accctccacg aagaggaaga agccgcgggg gttcctgctc agcaggcgca
      601 gggcagcctc tgtcatctcc atcagggagg ggtccagtgt ggagtctcgg tggatctcgt
      661 atttcatgtc tccaggctca aagagaccca tgagatgggt cacagacggg tccagggaag
      721 cctgcatgag ctcagtgcgg ttccacacat accgggcacc ctggcgcttc gccagccatt
      781 cctgcaccag attcttcccg tccagcctgg tcccaccttg gctgtagtca tctgggtact
      841 cagggtctgg ggttcccatg cgaaacatgt actttcggcc tccacctagg atcacgtcaa
      901 tgtccatgtt ggagatgagc tgcgtagcga tgtcctggca cccctcctgg cgggccgagg
      961 caggcacgtc ggcgtccgag taccagttgc ggttcaccgt gtgggcgtag gtgccggctg
     1021 gcgaggcgtg ctgcactcgt gtggtggtta ccactcccac tgacttccct gctttcttgg
     1081 cccgattcat cacggagatg acctcgttgc cgcgtgtcgt gttgcactgg ttaaagcggg
     1141 cggctgcact caagccaatg gtctggaagt tgcccttgac cccgcacagg taggccgtgg
     1201 ctgtggctcc actgtctggc acatgtttgt ctacattgta tgtcttggac agagccacat
     1261 atgggaagcg gtccatggcc aggggtatct caggccccag tttgtccttc ttctgccctt
     1321 ttaggatcct ggcagctgtc accgtagaca cccccatccc atcgcccagg aagatgatga
     1381 ggttcttggc ggctgtctgt gcaggctgca gcttcttggc ggcacccagg gcctcggctg
     1441 cctcgcggtt ccagaagtcc gggttctcct cctcaactgg gatgatgccc agggagagct
     1501 gtagcctcag gcccagcagc agcagcagca gcagcatgca gggccccagc atgtctggag
     1561 ggcagcagga cagtggcgag gcaggaatta gcttgaggtg cacaccaatg tggtgaatgg
     1621 tcaaatggcg tttattgtat cgagctaggc acttaaatac aatatctctg caatgcggaa
     1681 ttcgatatca agcttatcga tacggttcac taaacgagct ctgcttatat agacctccca
     1741 ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa
     1801 gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg
     1861 aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac
     1921 catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt
     1981 catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt
     2041 acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac
     2101 ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga
     2161 cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt
     2221 aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata
     2281 actagtcaat aatcaatgtc aacatggcgg taatgttgga catgagccaa tataaatgta
     2341 catattatga tatggataca acgtatgcaa tgggccaagc tcatggctga ctaatttttt
     2401 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     2461 gcttttttgg aggcctaggc ttttgcaaaa agctcgacgc gaggctcgag ggggggcccg
     2521 gtacccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt tttacaacgt
     2581 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     2641 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     2701 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggttacg
     2761 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     2821 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     2881 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     2941 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3001 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3061 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3121 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
     3181 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3241 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     3301 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3361 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     3421 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3481 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3541 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3601 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3661 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     3721 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3781 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3841 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     3901 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3961 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     4021 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     4081 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     4141 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     4201 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4261 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4321 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4381 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4441 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4501 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4561 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4621 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4681 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4741 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4801 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4861 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4921 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     4981 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     5041 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     5101 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     5161 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     5221 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     5281 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctcg
     5341 aaattaaccc tcactaaagg gaacaaaagc tggagctcca ccgcggtggc ggccgctcta
     5401 ggatcgatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     5461 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     5521 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     5581 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt
     5641 atgatcaatg aattgctcta gaggatcccc gggtacccac tgggctgtag gtggtggctg
     5701 gatggcctga gccccctcct ttcctggggt gcaggcagcc acagggcaca acctgcagtc
     5761 cctggggttg gggtagggag tccgagagca gccaaggggc aggtttggtg tcatgggctc
     5821 agtccccaga gcgcacgggg aggggaaggt tccatggcag gacctctgta tagggactcc
     5881 ggggatgacg actcgggctg gaggccaggc aggacgacag gaggtgggga gcaggagaac
     5941 tccgggatgg ggaagcagga gccccaggga cgggacactc agggagcagt ggccgtctcc
     6001 agcagcagca gggtcccggc cagcagagga agca
//
